Restriction endonuclease enhanced polymorphic sequence detection

ABSTRACT

Provided in part herein is an improved method for the detection of specific polymorphic alleles in a mixed DNA population. The method comprises enriching the relative percentage of a given polymorphic allele that is exponentially amplifiable by PCR. Provided also are methods for selectively enriching target nucleic acid, for example, fetal nucleic acid in a maternal sample. In the case of detecting fetal nucleic acid in a maternal sample, a restriction enzyme is introduced that can discriminate between the alleles of a polymorphic site. In some embodiments, the maternal allele is digested and nucleic acid comprising the paternal allele is relatively enriched.

RELATED PATENT APPLICATIONS

This patent application claims the benefit of U.S. Provisional PatentApplication No. 61/039,747, filed on Mar. 26, 2008, entitled RESTRICTIONENDONUCLEASE ENHANCED POLYMORPHIC SEQUENCE DETECTION and designated byattorney docket no. SEQ-6019-PV. This patent application also is relatedto U.S. Provisional Patent Application No. 60/908,167, filed on Mar. 26,2007 (designated by attorney docket no. SEQ-6008-PV), and PatentCooperation Treaty International Patent Application No.PCT/US2008/058317, filed on Mar. 26, 2008, and published as PublicationNo. WO2008/118988 on October 2, 2008 (designated by attorney docket no.SEQ-6008-PC), each entitled RESTRICTION ENDONUCLEASE ENHANCEDPOLYMORPHIC SEQUENCE DETECTION. The entirety of each of these threepatent applications is hereby incorporated herein by reference.

FIELD OF THE INVENTION

The invention in part pertains to methods for detecting specific allelesin a mixed nucleic acid sample. Methods provided herein can be used todetect the presence or absence of fetal nucleic acid in a maternalsample.

BACKGROUND

The analysis of circulating nucleic acids has revealed applications inthe non-invasive diagnosis, monitoring, and prognostication of manyclinical conditions. For example, for prenatal applications, circulatingfetal-specific sequences have been detected and constitute a fraction ofthe total DNA in maternal plasma. The diagnostic reliability ofcirculating DNA analysis depends on the fractional concentration of thetargeted sequence, the analytical sensitivity, and the specificity. Therobust discrimination of sequence differences (e.g., single-nucleotidepolymorphisms, or SNPs) between circulating DNA species is technicallychallenging and demands the adoption of highly sensitive and specificanalytical methods.

Current techniques to detect sequence differences in a DNA sampleinclude allele-specific PCR, restriction digest and Southern blothybridization, restriction endonuclease-mediated selective-PCR(REMS-PCR), and competitive PCR methods involving the use of fluorescentdetection probes.

Currently available techniques present several disadvantages. Forallele-specific PCR, it is often difficult to design assays with a highdegree of allele specificity (Nasis et al. Clin Chem. 2004April;50(4):694-701). Restriction digest/Southern blot methods requirehigher amounts of DNA template than the method provided herein, and lackthe sensitivity to detect polymorphic sequences comprising a lowrelative proportion of total DNA. Restriction endonuclease-mediatedselective-PCR (REMS-PCR) has the drawback of requiring a thermostablerestriction enzyme that cleaves the wild-type allele. REMS-PCR isdescribed in U.S. Pat. No. 6,261,768, which is hereby incorporated byreference. Use of the technique may not always be possible, and thisrequirement limits the general utility of the REMS-PCR approach.Competitive PCR lacks the sensitivity to detect polymorphic sequencescomprising a low relative proportion (<5%) of total DNA. Competitive PCRwith allele-specific fluorescent probes lacks the ability to multiplexassays higher than 2-3 assays in a single tube format. In addition,similar methods utilizing methylation differences between DNA species(for example, US Patent Application Publication No. 20070059707,entitled, “Methods for prenatal diagnosis of chromosomal abnormalities”,which is hereby incorporated by reference) are not effective at low copynumbers of genomic DNA.

SUMMARY

The invention in part provides sequence-specific cleavage of nucleicacid to selectively enrich for a particular target nucleic acid.Polymorphic loci are chosen such that only one allele at the polymorphiclocus is cleaved by a given cleavage agent, such as a restrictionendonuclease. Oligonucleotide primer pairs designed to flank thepolymorphism allow amplification of the polymorphic region, or amplicon,by amplification (e.g., PCR). Prior to or during amplification, nucleicacid samples are incubated with the given restriction endonuclease. Insome embodiments, the cleavage agent is introduced prior toamplification. This approach results in cleavage of the polymorphicallele or sequence comprising the polymorphic allele that is recognizedby the restriction endonuclease, if this allele is present. Cleavage ofany template nucleic acid within the amplicon sequence (i.e., betweenprimer pairs) prevents PCR amplification of this template. Therefore, ifonly one allele of a polymorphism is recognized by the cleavage agentand the corresponding nucleic acid sequence is cleaved by therestriction endonuclease, the relative percentage of the amplifiablealternate polymorphic allele is increased in a manner dependent on theefficiency and specificity of the restriction endonuclease activity.After amplification, the amplified polymorphic alleles can be genotypedor otherwise detected or discriminated by any method known in the art(e.g., using Sequenom's MassARRAY® technology or by RT-PCR).

In some embodiments, the invention in part provides a method fordetecting the presence or absence of a target allele at a polymorphiclocus in a sample, where the sample contains nucleic acid, whichcomprises: cleaving a nucleic acid comprising a non-target allele at ornear the polymorphic locus with a cleavage agent that recognizes andcleaves the non-target allele, but not the target allele; amplifyinguncleaved nucleic acid but not cleaved nucleic acid; and analyzing theamplification products from the previous step to determine the presenceor absence of the target allele. In certain embodiments, the method alsocomprises first obtaining a sample suspected of comprising nucleic acidwith target and non-target alleles. In some embodiments, the method isused to distinguish between two individuals, for example, between amother and a fetus, where the sample comprises both maternal and fetalnucleic acid. Optionally, the method may be used to quantify the targetnucleic acid relative to the non-target nucleic acid.

The invention also in part provides methods for enriching for targetnucleic acid, comprising cleaving nucleic acid comprising a non-targetallele with a restriction endonuclease that recognizes the nucleic acidcomprising the non-target allele but not the target allele; andamplifying uncleaved nucleic acid but not cleaved nucleic acid, wherethe uncleaved, amplified nucleic acid represents enriched target nucleicacid relative to non-target nucleic acid. In some embodiments, methodsprovided herein may be utilized to determine the presence or absence oftarget nucleic acid in a background of non-target nucleic acid. Incertain embodiments, the amplification products can be analyzed todiagnose, monitor or prognose a clinical condition. Likewise, theamplification products can be analyzed to assist in the diagnosis,prognosis or monitoring of a clinical condition or chromosomalabnormality. Nucleic acid may be selected such that it comprises anallele having a polymorphic site that is susceptible to selectivedigestion by a cleavage agent, for example.

Methods provided herein are useful for analyzing nucleic acid including,but not limited to, DNA, RNA, mRNA, oligonucleosomal, mitochondrial,epigenetically-modified, single-stranded, double-stranded, circular,plasmid, cosmid, yeast artificial chromosomes, artificial or man-madeDNA, including unique DNA sequences, and DNA that has been reversetranscribed from an RNA sample, such as cDNA, and combinations thereof.In some embodiments, methods provided herein are used to detect orselectively enrich RNA.

A nucleic acid may also be characterized as target nucleic acid ornon-target nucleic acid, where target nucleic comprises the targetallele and non-target nucleic acid comprises the non-target allele. Insome embodiments, the target nucleic acid comprises the paternal alleleand the non-target nucleic acid comprises the maternal allele. Incertain embodiments, the nucleic acid is cell-free nucleic acid orpartially cell-free nucleic acid. In some embodiments, the targetnucleic acid is apoptotic or partially apoptotic. In certainembodiments, the target nucleic acid is less than 2000, 1200, 1100,1000, 900, 800, 700, 600, 500, 400, 300, 200, 100, 80, 70, 60, 50, 40 orless base pairs in length.

Methods provided herein may be used to detect target nucleic acid in abiological sample. In some embodiments, the biological sample is from ananimal, often a human. In certain embodiments, the biological sample isselected from the group of whole blood, serum, plasma, umbilical cordblood, chorionic villi, amniotic fluid, cerbrospinal fluid, spinalfluid, lavage fluid, biopsy sample, urine, feces, sputum, saliva, nasalmucous, prostate fluid, semen, lymphatic fluid, bile, tears, sweat,breast milk, breast fluid, embryonic cells and fetal cells, and mixturethereof. In some embodiments, the sample is from a crime scene (e.g.,used for forensic analysis). In certain embodiments, the biologicalsample is obtained through non-invasive means, for example, a blood drawfrom a pregnant female. In another some embodiments, the biologicalsample is cell-free. In certain embodiments, the sample is a previouslyisolated sample of nucleic acids.

In some embodiments, the invention in part provides a method fordetecting the presence or absence of fetal nucleic acid in a maternalsample, where the sample contains nucleic acid, which comprises:cleaving nucleic acid comprising a maternal allele with a restrictionendonuclease that recognizes and cleaves the nucleic acid comprising thematernal allele but not the paternal allele; amplifying uncleavednucleic acid but not cleaved nucleic acid; and analyzing theamplification products from the previous step to determine the presenceor absence of fetal nucleic acid. In certain embodiments, the samplecomprises a mixture of nucleic acids. For example, the mixture maycomprise nucleic acid from different species or from differentindividuals. In some embodiments, the sample is from a pregnant female.Samples can be collected from human females at 1-4, 4-8, 8-12, 12-16,16-20, 20-24, 24-28, 28-32, 32-36, 36-40, or 40-44 weeks of fetalgestation, and sometimes between 5-28 weeks of fetal gestation. Incertain embodiments, methods provided herein may be used to detect thepresence or absence of fetal Y-chromosome nucleic acid, therebydetermining the sex of the fetus.

In some embodiments, the target nucleic acid comprises a paternalallele. In certain embodiments, the mother is homozygous at thepolymorphic site and the fetus is heterozygous at the polymorphic site.In the case when the mother is homozygous at the polymorphic site andthe fetus is heterozygous at the polymorphic site, the polymorphic siteis considered informative (e.g., see FIG. 5A for examples of informativeand non-informative cases). In certain embodiments, the maternalgenotype is determined in conjunction with methods provided herein. Insome embodiments, the mother is first genotyped (for example, usingperipheral blood mononuclear cells (PBMC) from a maternal whole bloodsample) to determine the non-target allele that will be recognized andcleaved by the cleavage agent. When the method is used for forensicpurposes, the victim may be first genotyped to determine the non-targetallele that will be recognized and cleaved by the cleavage agent.Likewise, when used for organ transplant-related applications, thetransplant recipient may be first genotyped to determine the non-targetallele that will be recognized and cleaved by the cleavage agent.

In certain embodiments, the sample contains nucleic acid from twodifferent individuals. Such instances include, but are not limited to,organ transplant recipients, transfusion recipients, and forensicapplications.

In certain embodiments, the sample is from an individual suspected ofsuffering from a disease, and the non-target allele is a wild-typeallele that is selectively cleaved in order to enrich for adisease-related point mutation. In certain embodiments, the disease iscancer. The ras proto-oncogenes, K-ras, N-ras, and H-ras, and the p53tumor suppressor gene are examples of genes which are frequently mutatedin human cancers. Specific mutations in these genes leads to activationor increased transforming potential.

The invention also in part provides methods useful for detecting rarealleles or low copy number alleles. In some embodiments, the targetallele is undetectable by conventional or unmodified genotyping methodsif the non-target allele is not selectively cleaved. In certainembodiments, the target allele is not detectable unless it isselectively enriched, for example, by methods provided herein. Incertain embodiments, the target allele concentration (e.g., alleleconcentration in a sample) is about 0.1% to about 40%, e.g., about 0.5%,1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%,15%,16%,17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%,31%, 32%, 33%, 34% or 35%, of total nucleic acid (e.g., total nucleicacid (e.g., total nucleic acid in a composition or sample), or is lessthan one of the foregoing percentages. Total nucleic acid includesmaternal nucleic acid and any fetal nucleic acid, and total nucleic acidincludes non-target allele and any target allele. When fetal nucleicacid is present, target allele is about 50% of the fetal nucleic acid,and non-target allele often includes the other about 50% of the fetalnucleic acid and all maternal nucleic acid, in some embodiments. Incertain embodiments, the target nucleic acid number is about 1 to about5,000 molecules, e.g., about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,17, 18, 19, 20, 25, 30, 35, 40, 50, 55, 60, 70, 80, 90, 100, 150, 200,250, 300, 400, 500, 600, 700, 800, 900 or 1000 molecules, or is lessthan one of the foregoing numbers of molecules. In certain embodiments,the target allele is a mutation, and the non-target allele is thewild-type allele. In certain embodiments, the target allele may beeither a somatic or germline mutation. In certain embodiments, anotherallele or sequence identifier in the same amplicon as the polymorphiclocus may be detected. For example, a sequence comprising a targetallele may be selectively enriched using methods provided herein, andanother sequence identifier may be detected by any method known in theart.

In certain embodiments, there are no other polymorphic loci within theamplicon that may be recognized by the cleavage agent. For example,there is only one polymorphic locus in the amplicon recognized by thecleavage agent in some embodiments.

In certain embodiments, the method optionally comprises first isolatingnucleic acid from the sample. DNA isolation from blood, plasma, or serumof the pregnant mother can be performed using any method known to oneskilled in the art. Any standard DNA isolation technique can be used toisolate the fetal DNA and the maternal DNA including, but not limitedto, QIAamp DNA Blood Midi Kit supplied by QIAGEN. Other standard methodsof DNA isolation are described, for example, in (Sambrook et al.,Molecular Biology: A laboratory Approach, Cold Spring Harbor, N. Y.1989; Ausubel, et al., Current protocols in Molecular Biology, GreenePublishing, Y, 1995). A method for isolation of plasma DNA is describedin Chiu et al., 2001, Clin. Chem. 47: 1607-1613, which is hereinincorporated by reference in its entirety. Other suitable methods areprovided in Example 2 of PCT International Application PublicationNumber 2007/028155, filed on Sep. 1, 2006.

Methods described herein allow for the use of any cleavage agent capableof distinguishing between two different sequences, and cleavingsomewhere within the amplicon sequence thereby preventing amplificationof the cleaved sequence. The difference between the sequences may be theresult of different alleles at one or more polymorphic sites within thesequence. In another example, the difference between the sequences maybe the result of two homologous sequences, for example, betweenparalogous genes or between highly homologous genes such as the RhDgene, which encodes the D polypeptide, and the RHCE gene, which encodesthe CcEe polypeptide. An example of a cleavage agent is a restrictionenzyme, also referred to as a restriction endonuclease. Multiplerestriction endonucleases (available from various vendors) may beselected that correspond to appropriate sequence differences. In someembodiments, the restriction enzyme is a thermostable restrictionenzyme. In certain embodiments, the restriction enzyme is Tsp509I. Incertain embodiments, a step is added to end the cleaving activity of thecleavage agent, for example, by introducing a protease and/or hightemperature prior to amplification.

A restriction endonuclease may be added prior to or duringamplification, for example, during an incubation step. In someembodiments, the restriction endonuclease is added less than 5 minutes,5 minutes, 10 minutes, 20 minutes, 30 minutes, 40 minutes, 50 minutes,60 minutes, 90 minutes or 120 or more minutes before amplification.Incubation time may be shortened if additional units of restrictionenzyme are added to the reaction. Conversely, longer incubation timesare often used to allow a reaction to proceed to completion with fewerunits of enzyme. This is contingent on how long a particular enzyme cansurvive (maintain activity) in a reaction. Some enzymes survive for longperiods (>16 hours) while others survive only an hour or less in areaction. In certain embodiments, the restriction enzyme digests greaterthan 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98% or 99% of the non-target nucleic acid. However, if digestion ofnon-target nucleic acid of less than 40% allows for useful enrichment oftarget nucleic acid, it is within the scope of the invention. In certainembodiments, the restriction enzyme digests substantially all of thenon-target nucleic acid. In certain embodiments, the restrictionendonuclease is a thermostable restriction endonuclease. Examples ofthermostable endonucleases include, but are not limited to, Bst NI, BslI, Tru 9I and Tsp 509 I. In certain embodiments, the cleavage agent isnot thermostable, especially when the digestion occurs prior to theamplification step. In some embodiments, the cleavage agent isthermostable and a majority of the digestion of the non-target nucleicacid occurs prior to the amplification step during a pre-incubationstep. In certain embodiments, the restriction enzyme digests greaterthan 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98% or 99% of the non-target nucleic acid prior to amplification.In another embodiment, one or more incubation steps may be introducedduring thermal cycling. Incubation steps are ideally at the optimaltemperature for digestion to occur. For example, for Tsp509I theincubation temperature may be 65 degrees C. In certain embodiments, astep is introduced to prevent or to reduce digestion during theamplification step, for example, by introducing a protease to disable acleavage agent that is a protein.

In some embodiments, the units of restriction enzyme added to the sampleis 0.10, 0.25, 0.50, 0.75, 1.0, 2.0 or more. Note that DNA substratesare digested at varying rates, therefore, the actual number of unitsrequired for a complete or substantially complete digestion may varyfrom assay to assay.

In certain embodiments, only one restriction endonuclease is used todigest one or more non-target alleles in a single reaction. For example,a multiplexed assay may be designed where a single restrictionendonuclease performs multiple (e.g., greater than 5, 10, 15, 20, 25,50, 100) digestions across the genome. In certain embodiments, more thanone restriction endonuclease (e.g., greater than or equal to 2, 3, 4, 5,6, 7, 8, 9, 10) is used to make multiple (e.g., greater than 5, 10, 15,20, 25, 50, 100) digestions across the genome.

Amplification may be performed after or during the cleavage of thenon-target allele, and prior to the detection of the target allele. Insome embodiments, amplification is performed after cleavage of thenon-target allele. Amplification can be performed by any method known inthe art, including but not limited to polymerase chain reaction (PCR),ligase chain reaction, transcription-based amplification, restrictionamplification, or rolling circle amplification, using primers thatanneal to the selected fetal DNA regions. Oligonucleotide primers areselected such that they anneal to the sequence to be amplified. In someembodiments, primers are designed such that one or both primers of theprimer pair contain sequence recognizable by one or more restrictionendonucleases.

Following amplification, the relative enrichment of the target allele inthe sample allows accurate detection of allele frequencies usingpractically any method of nucleic acid detection known in the art. Forexample, any of the following methods may be used, including, but notlimited to, primer extension or microsequencing methods, ligase sequencedetermination methods, mismatch sequence determination methods,microarray sequence determination methods, restriction fragment lengthpolymorphism (RFLP) procedures, PCR-based assays (e.g., TAQMAN® PCRSystem (Applied Biosystems)), nucleotide sequencing methods,hybridization methods, conventional dot blot analyses, single strandconformational polymorphism analysis (SSCP), denaturing gradient gelelectrophoresis (DGGE), heteroduplex analysis, mismatch cleavagedetection, detection by mass spectrometry, real time-PCR andpyrosequencing.

Methods provided herein may also be multiplexed at high levels in asingle reaction. For example, one or more alleles can be detectedsimultaneously. Multiplexing embodiments are particularly important whenthe genotype at a polymorphic locus is not known. In some instances, forexample when the mother is heterozygous at the polymorphic locus, theassay may not be informative. See FIG. 5A, which further describes theuse of polymorphic variants to detect fetal nucleic acid from a maternalsample. In some embodiments, 1 to 1,000 target alleles are assayed(e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54,55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72,73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 110, 120, 130, 140, 150, 160,170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300,310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440,450, 460, 470, 480, 490 or 500 target alleles are assayed), or a numberof target alleles more than one of the foregoing number of targetalleles is assayed, where each of the target alleles assayed may or maynot be informative (e.g., not every target allele is informative). Incertain embodiments, the genotype at the polymorphic locus is known. Incertain embodiments, 5 or more, 10 or more, 15 or more, 20 or more, 25or more, 30 or more, 35 or more, 40 or more, 45 or more, 50 or more, 55or more, 60 or more, 65 or more, 70 or more, 75 or more, 80 or more, 85or more or 90 or more target alleles are assayed (e.g., informativetarget alleles are assayed). The invention in part also includescombinations of different multiplex schemes provided herein.

In certain embodiments, the invention in part provides a method forquantifying a target allele at a polymorphic locus in a sample, wherethe sample contains nucleic acid, that comprises: digesting nucleic acidcontaining a maternal allele at the polymorphic locus with an enzyme,such as a restriction endonuclease, that selectively digests thematernal allele, where the selective digestion yields a DNA sampleenriched for fetal DNA; determining the maternal or paternal allelefrequency using polymorphic markers within the amplicon, and comparingthe paternal or maternal allele frequency to a control DNA sample. Insome embodiments, a difference in allele frequency is indicative of achromosomal abnormality. In certain embodiments, the control DNA sampleis a competitor oligonucleotide that is introduced to the assay in knownquantities.

In certain embodiments, the present invention provides a kit fordetecting the presence or absence of target nucleic acid. One componentof the kit is primers for amplifying the region of interest. Anothercomponent of the kit comprises probes for discriminating between thedifferent alleles of each nucleic acid species.

Certain non-limiting embodiments of the invention are further describedin the following Brief Description of the Drawings, Detailed Descriptionand claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is the HpyCH4V digest, which shows allele peak area ratios in aDNA mixture series. Peak area ratio is determined by dividing thecalculated peak area of the SNP allele not recognized by HpyCH4V (i.e.,target allele) by the total peak area of both SNP alleles present in themass spectrum.

FIG. 2 is the NlaIII digest, which shows allele peak area ratios in aDNA mixture series. Peak area ratio is determined by dividing thecalculated peak area of the SNP allele not recognized by NlaIII (i.e.,target allele) by the total peak area of both SNP alleles present in themass spectrum.

FIG. 3 is the HpyCH4V screenshots of 2% heterozygous DNA mixture. Notethe appearance of the ‘A’ and ‘T’ alleles after HpyCH4V digestion of theDNA samples for rs4329520 and rs4658481, respectively.

FIG. 4 is the NlaIII screenshots of 2% heterozygous DNA mixture. Notethe appearance of the ‘T’ and ‘A’ alleles after NlaIII digestion of theDNA samples for rs2050927 and rs4329520, respectively.

FIG. 5A shows the use of single nucleotide polymorphisms (SNP's) FetalIdentifiers to confirm the presence of fetal DNA by paternally-inheritedalleles. FIG. 5B shows representative mass spectra demonstrating thecorrelation between fetal DNA amounts estimated from AMG XY and fromFetal Identifier assays. The results were generated using the AMGprimers provided in FIG. 9A-9C.

FIG. 6 depicts typical performance results for a qualified fetalidentifier. Here the ability of the SNP assay to estimate the quantityof fetal DNA in the background of maternal DNA was verified for a totalof 1700 copies and a total of 170 copies using genomic DNA mixtures.Note that the standard deviation of the estimate of fetal DNA increasesdue to the significant influence of the sampling error at low copynumbers.

FIG. 7 shows the performance of multiplexed SNP assays (21 assays total)for detection of paternally-inherited alleles in a model system.

FIGS. 8A-8C provide the location design of the AMG primers. Theamplification primers are underlined once and the extend primers areunderlined twice. In addition, competitor sequences are provided.Competitor sequences may be used for quantitative methods. FIG. 8Cincludes a Results Table that shows the different masses generated byeach of the AMG and SRY assays, which may be used to interpret theresults from the assays.

FIG. 9 provides the location design of the albumin (ALB) primers. Theamplification primers are highlighted and the extend primer isunderlined twice. Where the PCR primers are provided alone, thesequence-specific portion of the primer is underlined, and the multiplextag is not underlined. In addition, competitor sequences are provided.Competitor sequences may be used for quantitative methods.

FIG. 10 shows the number of SNPs for the indicated Tsp509I digestedsample with greater than 15% primer extension rate and 0.4 or higherincrease in informative allele peak area ratio when compared to thematching undigested maternal DNA only (for mixtures) or undigestedmaternal PBMC DNA (for PBMC and plasma DNAs).

FIG. 11 shows results from 92 fetal identifiers tested in 117 plasmasamples from pregnant and non-pregnant women. The x-axis of the dot plotin the top portion indicates the number of fetal identifier allelesdetected in a plasma DNA sample (i.e., the number of informative SNPs).Each dot in the dot plot field represents a sample. The top portion ofthe panel comprises 27 non-pregnant plasma samples. The bottom portionof the panel comprises 90 pregnant, maternal plasma samples. The legendprovides sample type and fetal sex (if known).

FIG. 12 is a graph showing the probability of the number of informativeSNPs for each of the selected thresholds (1-6) at increasing numbers oftotal SNPs assayed.

DETAILED DESCRIPTION

It has been determined in the fields of biology and diagnostics thatcertain nucleic acids are present at very low concentrations in humans.In particular, fetal DNA has been found to exist in maternal plasma (Loet al. Lancet. Aug. 16, 1997;350(9076):485-7). This discovery hasfacilitated the development of non-invasive prenatal diagnosticapproaches based simply on the analysis of a maternal blood sample (Loet al. Am J Hum Genet. 1998 April;62(4):768-75). The non-invasive natureof maternal plasma-based approaches represents a major advantage overconventional methods of prenatal diagnosis, such as amniocentesis andchorionic villus sampling, which are associated with a small but finiterisk of fetal loss. However, a technical challenge experienced by manyworkers in the field relates to the ability to discriminate therelatively small amount of fetal DNA from the coexisting background ofmaternal DNA in maternal plasma. During pregnancy, fetal DNA amounts toapproximately 3-6% of the total DNA in maternal plasma. Hence, thediagnostic reliability of fetal DNA analysis in maternal plasmagenerally has depended on the accurate detection of fetal-specificmarkers.

Methods described herein solve this problem by enriching, relatively,the amount of low copy number nucleic acid before detecting orquantifying the alleles present in the sample. In the case of prenataldiagnostics, the use of restriction endonuclease enhanced polymorphicsequence detection allows for the selective, sensitive detection offetal nucleic acid from maternal samples. The fetal DNA in the maternalplasma sample is selectively enriched before detecting the allelespresent in the maternal sample. To enrich for fetal DNA present inplasma of the mother to allow accurate detection of fetal allelespresent in the sample, methods provided herein allow for the cleavage ofmaternal nucleic acid or nucleic acid of maternal origin. Thus, thematernal DNA can be substantially reduced, masked, or destroyedcompletely, and the sample is left with DNA enriched for DNA of fetalorigin. The selective reduction of maternal DNA can be performed usingone or more enzymes, such as restriction endonucleases, whichselectively digest nucleic acids which comprise maternal alleles.

The term “sample” as used herein refers to a composition, specimen orculture (e.g., microbiological cultures) that includes nucleic acids.The term “sample” includes biological and environmental samples. Asample may include a specimen of synthetic origin. Biological samplesinclude whole blood, serum, plasma, umbilical cord blood, chorionicvilli, amniotic fluid, cerbrospinal fluid, spinal fluid, lavage fluid(e.g., bronchoalveolar, gastric, peritoneal, ductal, ear, athroscopic),biopsy sample, urine, feces, sputum, saliva, nasal mucous, prostatefluid, semen, lymphatic fluid, bile, tears, sweat, breast milk, breastfluid, embryonic cells and fetal cells. A biological sample can bematernal blood, including maternal plasma or serum. In somecircumstances, a biological sample is acellular. In other circumstances,a biological sample does contain cellular elements or cellular remnantsin maternal blood. In some embodiments, a nucleic acid sample is, or isobtained from, an extracellular or acellular composition (e.g., bloodplasma, blood serum, urine).

In some embodiments, a sample comprises a mixture of nucleic acids. Forexample, the mixture may comprise nucleic acid from different species orfrom different individuals. In some embodiments, a sample is from apregnant female or a female suspected of being pregnant. In certainembodiments, the sample is procured through non-invasive means (e.g., ablood draw). In some embodiments the sample is from any animal,including but not limited to, human, non-human, mammal, reptile, cattle,cat, dog, goat, swine, pig, monkey, ape, gorilla, bull, cow, bear,horse, sheep, poultry, mouse, rat, fish, dolphin, whale, and shark, orany animal or organism that may be tested for the presence of targetnucleic acid.

In some embodiments, the biological sample is blood, and sometimesplasma. As used herein, the term “blood” encompasses whole blood or anyfractions of blood, such as serum and plasma as conventionally defined.Blood plasma refers to the fraction of whole blood resulting fromcentrifugation of blood treated with anticoagulants. Blood serum refersto the watery portion of fluid remaining after a blood sample hascoagulated. Environmental samples include environmental material such assurface matter, soil, water, crime scene samples, and industrialsamples, as well as samples obtained from food and dairy processinginstruments, apparatus, equipment, utensils, disposable andnon-disposable items. These examples are not to be construed as limitingthe sample types applicable to the present invention.

The term “non-invasive” as used herein refers to a method for collectinga sample that poses minimal risk to an individual (e.g., the mother,fetus, victim, etc.). An example of a non-invasive method is a blooddraw; whereas examples of invasive methods include amniocentesis andchorionic villus sampling, both of which constitute a finite risk to thefetus.

The terms “target” or “target nucleic acid” as used herein refer to anymolecule whose presence is to be detected or measured or whose function,interactions or properties are to be studied, where target nucleiccomprises the target allele and non-target nucleic acid comprises thenon-target allele. Fetal nucleic acid may comprise both target nucleicacid and non-target nucleic when the fetus is heterozygous at apolymorphic locus. Other examples of target nucleic acid include, butare not limited to, trace nucleic acid, mutated nucleic acid, viralnucleic acid and transplant nucleic acid.

The terms “nucleic acid” and “nucleic acid molecule” may be usedinterchangeably herein. The terms refer to oligonucleotides, oligos,polynucleotides, deoxyribonucleotide (DNA), genomic DNA, mitochondrialDNA (mtDNA), complementary DNA (cDNA), bacterial DNA, viral DNA, viralRNA, RNA, message RNA (mRNA), transfer RNA (tRNA), ribosomal RNA (rRNA),siRNA, catalytic RNA, clones, plasmids, M13, P1, cosmid, bacteriaartificial chromosome (BAC), yeast artificial chromosome (YAC),amplified nucleic acid, amplicon, PCR product and other types ofamplified nucleic acid, RNA/DNA hybrids and polyamide nucleic acids(PNAs), all of which can be in either single- or double-stranded form,and unless otherwise limited, would encompass known analogs of naturalnucleotides that can function in a similar manner as naturally occurringnucleotides and combinations and/or mixtures thereof. Thus, the term“nucleotides” refers to both naturally-occurring andmodified/non-naturally-occurring nucleotides, including nucleoside tri,di, and monophosphates as well as monophosphate monomers present withinpolynucleic acid or oligonucleotide. A nucleotide may also be a ribo;2′-deoxy; 2′,3′-deoxy as well as a vast array of other nucleotide mimicsthat are well-known in the art. Mimics include chain-terminatingnucleotides, such as 3′-O-methyl, halogenated base or sugarsubstitutions; alternative sugar structures including nonsugar, alkylring structures; alternative bases including inosine; deaza-modified;chi, and psi, linker-modified; mass label-modified; phosphodiestermodifications or replacements including phosphorothioate,methylphosphonate, boranophosphate, amide, ester, ether; and a basic orcomplete internucleotide replacements, including cleavage linkages sucha photocleavable nitrophenyl moieties.

In the case of RNA, an RNA may be placentally-expressed RNA in maternalplasma. Background maternal RNA may be selectively digested according tomethods provided herein. Also, methods herein may further comprise anadditional step of discriminating alleles of RNA which involves reversetranscriptase polymerase chain reaction (RT-PCR). In certainembodiments, fetal RNA may be extracted from maternal body fluids,sometimes whole blood, and sometimes plasma or serum using e. g. RNAextraction methods such as, but not limited to, gelatin extractionmethod; silica, glass bead, or diatom extraction method; guanidiniumthiocyanate acid-phenol based extraction methods; guanidiniumthiocyanate acid based extraction methods; guanidine-hydrochloride basedextraction methods; methods using centrifugation through cesium chlorideor similar gradients; phenol-chloroform based extraction methods; and/orother available RNA extraction methods, as are known in the art for usein extraction of intracellular RNA, including commercially available RNAextraction methods, e. g. by using or adapting or modifying methods ofBoom et al. (1990, J. Clin. Microbiol. 28: 495-503); Cheung et al.(1994, J. Clin. Microbiol. 32: 2593-2597); Boom et al. (1991, J. Clin.Microbiol. 29: 1804-1811); Chomczynski and Sacchi (1987, AnalyticalBiochem. 162: 156-159); Chomczynski, (1993, Biotech. 15: 532-537);Chomczynski and Mackey (1995, Biotechniques 19: 942-945); Chomczynskiand Mackey (1995, Anal. Biochem. 225: 163-164); Chirgwin et al. (1979,Biochem. 18: 5294-5299); Fournie et al. (1986 Anal. Biochem. 158:250-256); and WO97/35589.

The term “amplification reaction” as used herein refers to any in vitromeans for multiplying the copies of nucleic acid. “Amplifying” as usedherein refers to a step of submitting a sample to conditions sufficientto allow for amplification. Components of an amplification reaction mayinclude, but are not limited to, e.g., primers, a polynucleotidetemplate, polymerase, nucleotides, dNTPs and the like. The term“amplifying” typically refers to an “exponential” increase in targetnucleic acid. However, “amplifying” as used herein can also refer tolinear increases in the numbers of a select target sequence of nucleicacid, but is different than a one-time, single primer extension step.“Polymerase chain reaction” or “PCR” as used herein refers to a methodwhereby a specific segment or subsequence of a target double-strandedDNA, is amplified in a geometric progression. PCR is well known to thoseof skill in the art; see, e.g., U.S. Pat. Nos. 4,683,195 and 4,683,202;and PCR Protocols: A Guide to Methods and Applications, Innis et al.,eds, 1990.

“Oligonucleotide” as used herein refers to linear oligomers of naturalor modified nucleosidic monomers linked by phosphodiester bonds oranalogs thereof. Oligonucleotides include deoxyribonucleosides,ribonucleosides, anomeric forms thereof, peptide nucleic acids (PNAs),and the like, capable of specifically binding to a target nucleic acid.Usually monomers are linked by phosphodiester bonds or analogs thereofto form oligonucleotides ranging in size from a few monomeric units,e.g., 3-4, to several tens of monomeric units, e.g., 40-60. Whenever anoligonucleotide is represented by a sequence of letters, such as“ATGCCTG,” it will be understood that the nucleotides are in 5′-3′ orderfrom left to right and that “A” denotes deoxyadenosine, “C” denotesdeoxycytidine, “G” denotes deoxyguanosine, “T” denotes deoxythymidine,and “U” denotes the ribonucleoside, uridine, unless otherwise noted.Oligonucleotides often comprise the four natural deoxynucleotides;however, they may also comprise ribonucleosides or non-naturalnucleotide analogs. Where an enzyme has specific oligonucleotide orpolynucleotide substrate requirements for activity, e.g., singlestranded DNA, RNA/DNA duplex, or the like, then selection of appropriatecomposition for the oligonucleotide or polynucleotide substrates is wellwithin the knowledge of one of ordinary skill.

As used herein “oligonucleotide primer”, or simply “primer”, refers to apolynucleotide sequence that hybridizes to a sequence on a nucleic acidtemplate and facilitates the amplification of the nucleic acid template,or otherwise plays a role in the detection of the nucleic acid molecule.In amplification embodiments, an oligonucleotide primer serves as apoint of initiation of nucleic acid synthesis. Primers can be of avariety of lengths and are often less than 50 nucleotides in length, forexample 12-25 nucleotides, in length. The length and sequences ofprimers for use in PCR can be designed based on principles known tothose of skill in the art.

The term “template” refers to any nucleic acid molecule that can be usedfor amplification in methods described herein. RNA or DNA that is notnaturally double stranded can be made into double stranded DNA so as tobe used as template DNA. Any double stranded DNA or preparationcontaining multiple, different double stranded DNA molecules can be usedas template DNA to amplify a locus or loci of interest contained in thetemplate DNA.

The term “amplicon” as used herein refers to amplified DNA that has been“copied” once or multiple times, e.g. by polymerase chain reaction. Theamplicon sequence falls between the amplification primers.

The term “polymorphic locus” as used herein refers to a nucleic acidregion that comprises a polymorphism. The nucleic acid region may be 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 35, 40, 50,60, 70, 80, 90, 100, 200, 300, 400, 500 or more nucleotides in length.The term “polymorphism” as used herein refers to an allelic variant.Polymorphisms can include single nucleotide polymorphisms (SNP's) aswell as simple sequence length polymorphisms, for example. Apolymorphism can be due to one or more nucleotide substitutions at oneallele in comparison to another allele or can be due to an insertion ordeletion, duplication, inversion and other alterations of one or morenucleotides. Certain polymorphisms include, but are not limited to,restriction fragment length polymorphisms (RFLPs), insertions/deletions,short tandem repeats, such as di-, tri-or tetra-nucleotide repeats(STRs), and the like. As used herein, the term “polymorphism” includesepigenetic variants, as long as cleavage by non-epigenetic-specificcleavage agents is utilized.

The term “allele” as used herein is one of several alternate forms of agene or non-coding regions of DNA that occupy the same position on achromosome. The term allele can be used to describe DNA from anyorganism including but not limited to bacteria, viruses, fungi,protozoa, molds, yeasts, plants, humans, non-humans, animals, andarcheabacteria.

Alleles can have an identical sequence or can vary by a singlenucleotide or more than one nucleotide. With regard to organisms thathave two copies of each chromosome, if both chromosomes have the sameallele, the condition is referred to as homozygous. If the alleles atthe two chromosomes are different, the condition is referred to asheterozygous. For example, if the locus of interest is SNP X onchromosome 1, and the maternal chromosome contains an adenine at SNP X(A allele) and the paternal chromosome contains a guanine at SNP X (Gallele), the individual is heterozygous A/G at SNP X.

As used herein, the term “mutant alleles” may refer to variant allelesthat are associated with a disease state, e.g., cancer. The term“sequence identifier” as used herein refers to any sequence differencethat exists between two sequences that can be used to differentiate thesequences. In some embodiments, the sequence identifier does not includemethylation differences.

As used herein, the term “genotype” refers to the identity of thealleles or non-homologous variants present in an individual or sample.The term “genotyping a sample” or “genotyping an individual” refers todetermining a specific allele or specific nucleotide(s) orpolymorphism(s) in a sample or carried by an individual at particularregion(s).

The term “selectively” as used herein does not suggest an absoluteevent, but instead a preferential event. For example, “selectivelycleaved” is used to indicate one sequence (for example, the non-targetsequence) is preferentially cleaved or digested over another sequence(for example, the target sequence). However, some of a target sequencemay also be cleaved due to a lack of specificity with the cleavage agentor other variables introduced during the cleavage process.

The term “cleavage agent” as used herein refers to any means that iscapable of differentially cleaving two or more sequences based on asequence difference that exists between the two or more sequences. Thecleavage agent may be an enzyme in some embodiments. The cleavage agentmay be natural, synthetic, unmodified or modified. In some embodiments,the cleavage agent is a restriction endonuclease. Restrictionendonucleases, alternatively called restriction enzymes, are a class ofbacterial enzymes that cut or digest DNA at specific sites. Type Irestriction endonucleases occur as a complex with the methylase and apolypeptide that binds to the recognition site on DNA. They are oftennot very specific and cut at a remote site. Type II restrictionendonucleases are the classic experimental tools. They have veryspecific recognition and cutting sites. The recognition sites are short,4-8 nucleotides, and are usually palindromic sequences. Because bothstrands have the same sequence running in opposite directions theenzymes make double-stranded breaks, which, if the site of cleavage isoff-center, generates fragments with short single-stranded tails; thesecan hybridize to the tails of other fragments and are called stickyends. They are generally named according to the bacterium from whichthey were isolated (first letter of genus name and the first two lettersof the specific name). The bacterial strain is identified next andmultiple enzymes are given Roman numerals. For example the two enzymesisolated from the R strain of E. coli are designated Eco RI and Eco RII.In some embodiments, the restriction enzyme is a type II restrictionendonuclease. In another some embodiments, the restriction enzyme isthermostable.

The term “chromosomal abnormality” as used herein refers to a deviationbetween the structure of the subject chromosome and a normal homologouschromosome. The term “normal” refers to the predominate karyotype orbanding pattern found in healthy individuals of a particular species. Achromosomal abnormality can be numerical or structural, and includes butis not limited to aneuploidy, polyploidy, inversion, a trisomy, amonosomy, duplication, deletion, deletion of a part of a chromosome,addition, addition of a part of chromosome, insertion, a fragment of achromosome, a region of a chromosome, chromosomal rearrangement, andtranslocation. A chromosomal abnormality can be correlated with presenceof a pathological condition or with a predisposition to develop apathological condition.

Uses and Advantages Associated with Methods Described Herein

The invention in part provides nucleic acid-based assays that areparticularly useful for non-invasive prenatal testing. Methods providedherein may be used, inter alia, to determine the presence of fetalnucleic acid in a sample, to determine the amount of fetal nucleic acidin a sample, to determine the sex of a fetus, and to enrich for a targetnucleic acid sequence. The invention in part may be combined with otherprenatal methods, such as those described in U.S. application Ser. No.12/027,954, filed Feb. 7, 2008; PCT Application No. PCT/US07/69991,filed May 30, 2007; PCT Application No. PCT/US07/071232, filed Jun. 15,2007; PCT Patent Publication Numbers WO 2009/032779 and WO 2009/032781,both filed Aug. 28, 2008, PCT Patent Publication Number WO 2008/118988,filed Mar. 26, 2008, and PCT Patent Application Number PCT/EP05/012707,filed Nov. 28, 2005; or any of the prenatal diagnostic (both invasiveand non-invasive) methods disclosed in PCT Patent Publication No. WO2008/157264, filed on Jun. 12, 2008, all of which are herebyincorporated by reference.

The invention in part may be used to more accurately detect fetal DNAusing high frequency polymorphisms that match the criteria providedherein. These polymorphisms are alternatively called fetal identifiers.The criteria includes one or more of the following:

1) One allele of the SNP is recognized by the cleavage agent;

2) The alternate SNP allele is not recognized by the same cleavageagent;

3) No other sites for the cleavage are found ±50 base pair of the SNPwithin the PCR amplicon; and

4) (Optionally) The minor allele frequency is greater than 0.4(sometimes across a range of populations).

Examples of fetal identifiers are set forth in Table 16. In someembodiments, the method of detecting the presence or absence of fetalnucleic acid in a sample comprises obtaining or possessing a nucleicacid sample known to be of maternal origin and suspected of comprisingfetal nucleic acid; analyzing the nucleic acid sample to determine thematernal genotype at one or more nucleotide polymorphisms selected fromthe group consisting of the polymorphisms set forth in Table 16; andanalyzing the nucleic acid sample to determine the fetal genotype of oneor more nucleotide polymorphisms selected from the group consisting ofthe polymorphisms set forth in Table 16, where a fetal genotypepossessing a paternally-inherited allele indicates the presence of fetalnucleic acid, further where nucleic acid comprising a maternal allele isdigested using methods provided herein. In some embodiments, one or moreof the polymorphisms set forth in Table 16 are used in conjunction withmethods provided herein. In another some embodiments, one or more of themultiplex schemes provided in Table 11 is used according to methodsprovided herein. In certain embodiments, the maternal genotypes arefirst determined from DNA that is substantially free of fetal nucleicacid. For example, where the sample is blood of from blood, the maternalgenotypes may be determined from the portion of the blood that comprisesnucleated maternal cells (e.g., white blood cells). In some embodiments,the DNA that is substantially free of fetal nucleic acid is fromperipheral blood mononuclear cells. In certain embodiments, the amountof fetal DNA is determined by comparing the relative amount ofpaternally-inherited alleles to an internal control (e.g., competitoroligonucleotide).

In Table 11, each primer of the amplification primer pair may comprisethe entire sequence shown or only the non-underlined sequence, where theunderlined portion of the primer is a tag sequence (ACGTTGGATG) forimproved multiplexing and the non-underlined portion is asequence-specific primer sequence. The tag sequence may be any tagsequence known in the art that improves multiplexing. In certainembodiments, the invention in part includes primers that aresubstantially similar to the primers provided herein, for example, about90% or more identical (e.g., primers differ by 1, 2, 3, 4, 5, 6, 7, 8, 9or 10 nucleotide mismatches, or 1-3 nucleotide mismatches, when alignedwith one another), and further where the primers are still specific fora given nucleic acid region. For example, one or more bases of a primersequence may be changed or substituted, for example with an inosine, butthe primer still maintains the same specificity and plexing ability.Bases indicated by uppercase text are complementary to the nucleic acidsequence to which the primer hybridizes, and bases indicated bylowercase text are not complementary to the nucleic acid sequence towhich the primer hybridizes. Bases indicated in lower case text can beselected to shift or adjust the mass of primers and amplificationproducts.

In particular embodiments, a sequence tag is attached to a plurality ofprimary and secondary primer pairs provided in Table 11. The sequencetag can be attached to either one or both of the primary and secondaryprimers from each pair. Typically, the sequence tag is attached to theprimary and secondary primer of each pair. The sequence tags used hereincan range from 5 up to 20, from 5 up to 30, from 5 up to 40, or from 5up to 50 nucleotides in length, with a sequence tag of 10-mer lengthbeing particularly useful in methods provided herein. The sequence tagneed not be the same sequence for each primer pair in the multiplexedamplification reaction, nor the same sequence for a primary andsecondary primer within a particular amplification pair. In a particularembodiment, the sequence tag is the same for each primer in themultiplexed amplification reaction. For example, in certain embodiments,the sequence tag is a 10-mer, such as -ACGTTGGATG-, and is attached tothe 5′ end of each primary and secondary primer. In particularembodiments of methods provided herein, only a single primer pair isused to amplify each particular nucleic acid target-region.

In certain embodiments, methods provided herein may be used to improvethe detection the Y-chromosome in a maternal sample, which may be usedto determine the sex of a fetus. The presence or absence of theY-chromosome in a maternal sample may be determined by performing theSRY assay described in Example 3. The SRY assay is a highly sensitivequantitative internal standard assay that detects trace amounts of theY-chromosome. In certain embodiments, other polymorphisms located on theY-chromosome may be assayed according to methods provided herein.

The presence or absence of the Y-chromosome in a maternal sample mayalso be determined by performing the AMG assay provided herein. Thepresence or absence of a target nucleic acid may be determined incombination with other assays, such as an RhD assay, blood type assay orsex test assay. Methods provided herein may also be used for otherapplications, including but not limited to, paternity testing, forensicsor quality control assays.

In addition to prenatal applications, methods provided herein findutility in a range of applications, including, but not limited to,detecting rare cancer mutations, detecting transplant rejection andforensics.

In certain embodiments, the total copy number of nucleic acid moleculesfor the human serum albumin (ALB) gene is determined. Methods fordetermining the total copy number of nucleic acid present in a samplecomprise detecting albumin-specific extension products and comparing therelative amount of the extension products to competitors introduced tothe sample. In certain embodiments, the invention in part providescompositions and methods to determine the relative amount of fetal DNAin a sample (e.g., when the sample is plasma from a pregnant womancarrying a male fetus), which comprises annealing one or more albumingene sequences to the fetal DNA, the primers provided in FIG. 9;performing a primer extension reaction; and analyzing the primerextension products to determine the relative amount of ALB extensionproducts, where maternal albumin nucleic acid has been reduced usingmethods provided herein. In certain embodiments, the fetal ALB ampliconis first amplified using the amplification primers provided in FIG. 9.The assay is useful to measure how much nucleic acid (e.g., total copynumber) is present in a sample or loaded into a particular reaction. Theassay may serve as an internal control and a guide to the likelihood ofsuccess for a particular PCR reaction. For example, if only 400 copiesof ALB are measured then the probability of detecting any fetal DNA maybe considered low. In certain embodiments, the competitors provided inFIG. 9 are introduced as an internal standard to determine copy number.In some embodiments, 200, 300, 400, 500, 600, 700, 800 or morecompetitor molecules are introduced to the assay.

Methods described herein provide a number of advantages. Methodsprovided herein allow a high sensitivity to detect polymorphic alleles(e.g., fetal identifiers) present at low relative percentages in a DNAmixture and present at low copy number, for example. Methods providedherein may also be incorporated into multiplexed assays in a singlereaction in certain embodiments. Methods described herein are readilyimplemented, and only add a single additional step to the many currentdetection methods, for example.

Nucleases

Cleavage methods and procedures for selecting restriction enzymes forcutting nucleic acid at specific sites are well known to the skilledartisan. For example, many suppliers of restriction enzymes provideinformation on conditions and types of DNA sequences cut by specificrestriction enzymes, including New England BioLabs, Pro-Mega Biochems,Boehringer-Mannheim, and the like. Nucleic acid to be cleaved oftenis/are free of certain contaminants such as phenol, chloroform, alcohol,EDTA, detergents, or excessive salts, all of which can interfere withrestriction enzyme activity, in certain embodiments.

Embodiments of the invention can be assembled from multiple restrictionendonucleases (available from various vendors) that are chosen tocorrespond to appropriate polymorphic alleles, as long as a restrictionendonuclease selects for one polymorphic allele over another andperforms a digestion within the amplicon sequence such that it preventsa subsequent amplification event. In some embodiments, the amplicon ischosen such that it contains a variable nuclease restriction site andsequence identifier, which may or may not be the same as the restrictionsite. Also, the restriction enzyme need not cleave at the polymorphicsite, for example, at the variable nucleotide of a SNP.

Restriction enzymes are traditionally classified into three types on thebasis of subunit composition, cleavage position, sequence-specificityand cofactor-requirements. However, amino acid sequencing has uncoveredextraordinary variety among restriction enzymes and revealed that at themolecular level there are many more than three different kinds.

Type I enzymes are complex, multisubunit, combinationrestriction-and-modification enzymes that cut DNA at random far fromtheir recognition sequences. Originally thought to be rare, we now knowfrom the analysis of sequenced genomes that they are common. Type Ienzymes are of considerable biochemical interest but they have littlepractical value since they do not produce discrete restriction fragmentsor distinct gel-banding patterns.

Type II enzymes cut DNA at defined positions close to or within theirrecognition sequences. They produce discrete restriction fragments anddistinct gel banding patterns, and they are the only class used in thelaboratory for DNA analysis and gene cloning. Type II enzymes frequentlydiffer so utterly in amino acid sequence from one another, and indeedfrom every other known protein, that they likely arose independently inthe course of evolution rather than diverging from common ancestors.

The most common type II enzymes are those like HhaI, HindIII and NotIthat cleave DNA within their recognition sequences. Enzymes of this kindare available commercially. Most recognize DNA sequences that aresymmetric because they bind to DNA as homodimers, but a few, (e.g.,BbvCI: CCTCAGC) recognize asymmetric DNA sequences because they bind asheterodimers. Some enzymes recognize continuous sequences (e.g., EcoRI:GAATTC) in which the two half-sites of the recognition sequence areadjacent, while others recognize discontinuous sequences (e.g., BglI:GCCNNNNNGGC) in which the half-sites are separated. Cleavage leaves a3′-hydroxyl on one side of each cut and a 5′-phosphate on the other.They require only magnesium for activity and the correspondingmodification enzymes require only S-adenosylmethionine. They tend to besmall, with subunits in the 200-350 amino acid range.

The next most common type II enzymes, usually referred to as “type IIs”are those like FokI and AlwI that cleave outside of their recognitionsequence to one side. These enzymes are intermediate in size, 400-650amino acids in length, and they recognize sequences that are continuousand asymmetric. They comprise two distinct domains, one for DNA binding,the other for DNA cleavage. They are thought to bind to DNA as monomersfor the most part, but to cleave DNA cooperatively, through dimerizationof the cleavage domains of adjacent enzyme molecules. For this reason,some type IIs enzymes are much more active on DNA molecules that containmultiple recognition sites. A wide variety of Type IIS restrictionenzymes are known and such enzymes have been isolated from bacteria,phage, archeabacteria and viruses of eukaryotic algae and arecommercially available (Promega, Madison Wis.; New England Biolabs,Beverly, Mass.). Examples of Type IIS restriction enzymes that may beused with methods described herein include, but are not limited toenzymes such as those listed in Table IA.

Recognition/ Enzyme-Source Cleavage Site Supplier Alw I - Acinetobacterlwoffii GGATC(4/5) NE Biolabs Alw26 I - Acinetobacter lwoffi GTCTC(1/5)Promega Bbs I - Bacillus laterosporus GAAGAC(2/6) NE Biolabs Bbv I -Bacillus brevis GCAGC(8/12) NE Biolabs BceA I - Bacillus cereus 1315IACGGC(12/14) NE Biolabs Bmr I - Bacillus megaterium CTGGG(5/4) NEBiolabs Bsa I - Bacillus stearothermophilus 6-55 GGTCTC(1/5) NE BiolabsBst71 I - Bacillus stearothermophilus 71 GCAGC(8/12) Promega BsmA I -Bacillus stearothermophilus A664 GTCTC(1/5) NE Biolabs BsmB I - Bacillusstearothermophilus B61 CGTCTC(1/5) NE Biolabs BsmF I - Bacillusstearothermophilus F GGGAC(10/14) NE Biolabs BspM I - Bacillus species MACCTGC(4/8) NE Biolabs Ear I - Enterobacter aerogenes CTCTTC(1/4) NEBiolabs Fau I - Flavobacterium aquatile CCCGC(4/6) NE Biolabs Fok I -Flavobacterium okeonokoites GGATG(9/13) NE Biolabs Hga I - Haemophilusgallinarum GACGC(5/10) NE Biolabs Ple I - Pseudomonas lemoigneiGAGTC(4/5) NE Biolabs Sap I - Saccharopolyspora species GCTCTTC(1/4) NEBiolabs SfaN I - Streptococcus faecalis ND547 GCATC(5/9) NE BiolabsSth132 I - Streptococcus thermophilus CCCG(4/8) No commercial supplier(Gene ST132 195: 201-206 (1997))

A third major kind of type II enzyme, more properly referred to as “typeIV” are large, combination restriction-and-modification enzymes,850-1250 amino acids in length, in which the two enzymatic activitiesreside in the same protein chain. These enzymes cleave outside of theirrecognition sequences; those that recognize continuous sequences (e.g.,Eco57I: CTGAAG) cleave on just one side; those that recognizediscontinuous sequences (e.g., BcgI: CGANNNNNNTGC) cleave on both sidesreleasing a small fragment containing the recognition sequence. Theamino acid sequences of these enzymes are varied but their organizationare consistent. They comprise an N-terminal DNA-cleavage domain joinedto a DNA-modification domain and one or two DNA sequence-specificitydomains forming the C-terminus, or present as a separate subunit. Whenthese enzymes bind to their substrates, they switch into eitherrestriction mode to cleave the DNA, or modification mode to methylateit.

As discussed above, the length of restriction recognition sites varies.For example, the enzymes EcoRI, SacI and SstI each recognize a 6base-pair (bp) sequence of DNA, whereas NotI recognizes a sequence 8 bpin length, and the recognition site for Sau3AI is only 4 bp in length.Length of the recognition sequence dictates how frequently the enzymewill cut in a random sequence of DNA. Enzymes with a 6 bp recognitionsite will cut, on average, every 4⁶ or 4096 bp; a 4 bp recognition sitewill occur roughly every 256 bp.

Different restriction enzymes can have the same recognition site—suchenzymes are called isoschizomers. Table IB shows that the recognitionsites for SacI and SstI are identical. In some cases isoschizomers cutidentically within their recognition site, but sometimes they do not.Isoschizomers often have different optimum reaction conditions,stabilities and costs, which may influence the decision of which to use.Table IB is provided only to show exemplary restriction enzymes, anddoes not limit the scope of the invention in any way.

TABLE IB Enzyme Recognition Sequence BamH I GGATCC CCTAGG Not I GCGGCCGCCGCCGGCG Sau3A I GATC CTAG Sac I GAGCTC CTCGAG Sst I GAGCTC CTCGAG HinfI GANTC CTNAG Xho II PuGATCPy PyCTAGPu

Restriction recognition sites can be unambiguous or ambiguous. Theenzyme BamHI recognizes the sequence GGATCC and no others; therefore itis considered “unambiguous.” In contrast, HinfI recognizes a 5 bpsequence starting with GA, ending in TC, and having any base between (inTable IB, “N” stands for any nucleotide). HinfI has an ambiguousrecognition site. XhoII also has an ambiguous recognition site: Pystands for pyrimidine (T or C) and Pu for purine (A or G), so XhoII willrecognize and cut sequences of AGATCT, AGATCC, GGATCT and GGATCC.

The recognition site for one enzyme may contain the restriction site foranother. For example, note that a BamHI recognition site contains therecognition site for Sau3AI. Consequently, all BamHI sites will cut withSau3AI. Similarly, one of the four possible XhoII sites will also be arecognition site for BamHI and all four will cut with Sau3AI.

Also from Table IB, most recognition sequences are palindromes—they readthe same forward (5′ to 3′ on the top strand) and backward (5′ to 3′ onthe bottom strand). Most, but certainly not all recognition sites forcommonly-used restriction enzymes are palindromes. Most restrictionenzymes bind to their recognition site as dimers (pairs).

Nucleic Acid Detection

Whether detecting sequence differences, detecting amplification productsor primer extension products, any detection or discrimination methodknown may be utilized. These methods include, but are not limited to,primer extension reactions, mass spectrometry, hybridization using atleast one probe, hybridization using at least one fluorescently labeledprobe, direct sequencing, cloning and sequencing, and electrophoresis.Polymorphism detection methods known may also include, for example,microsequencing methods, ligase sequence determination methods (e.g.,U.S. Pat. Nos. 5,679,524 and 5,952,174, and WO 01/27326), digital PCR(U.S. Pat. No. 6,143,496), mismatch sequence determination methods(e.g., U.S. Pat. Nos. 5,851,770; 5,958,692; 6,110,684; and 6,183,958),microarray sequence determination methods, restriction fragment lengthpolymorphism (RFLP) procedures, PCR-based assays (e.g., TAQMAN® PCRSystem (Applied Biosystems)), nucleotide sequencing methods,hybridization methods, conventional dot blot analyses, single strandconformational polymorphism analysis (SSCP, e.g., U.S. Pat. Nos.5,891,625 and 6,013,499; Orita et al., Proc. Natl. Acad. Sci. U.S.A 86:27776-2770 (1989)), denaturing gradient gel electrophoresis (DGGE),heteroduplex analysis, mismatch cleavage detection, and techniquesdescribed in Sheffield et al., Proc. Natl. Acad. Sci. USA 49: 699-706(1991), White et al., Genomics 12: 301-306 (1992), Grompe et al., Proc.Natl. Acad. Sci. USA 86: 5855-5892 (1989), and Grompe, Nature Genetics5: 111-117 (1993), detection by mass spectrometry (e.g., US 20050079521,which is hereby incorporated by reference), real time-PCR (e.g., U.S.Pat. No. 5,210,015, U.S. Pat. No. 5,487,972, both of which are herebyincorporated by reference), or hybridization with a suitable nucleicacid primer specific for the sequence to be detected. Suitable nucleicacid primers can be provided in a format such as a gene chip.

Primer extension polymorphism detection methods, also referred to hereinas “microsequencing” methods, typically are carried out by hybridizing acomplementary oligonucleotide to a nucleic acid carrying the polymorphicsite. In these methods, the oligonucleotide typically hybridizesadjacent to the polymorphic site. As used herein, the term “adjacent”refers to the 3′ end of the extension oligonucleotide being sometimes 1nucleotide from the 5′ end of the polymorphic site, often 2 or 3, and attimes 4, 5, 6, 7, 8, 9, or 10 nucleotides from the 5′ end of thepolymorphic site, in the nucleic acid when the extension oligonucleotideis hybridized to the nucleic acid. The extension oligonucleotide then isextended by one or more nucleotides, often 1, 2, or 3 nucleotides, andthe number and/or type of nucleotides that are added to the extensionoligonucleotide determine which polymorphic variant or variants arepresent. Oligonucleotide extension methods are disclosed, for example,in U.S. Pat. Nos. 4,656,127; 4,851,331; 5,679,524; 5,834,189; 5,876,934;5,908,755; 5,912,118; 5,976,802; 5,981,186; 6,004,744; 6,013,431;6,017,702; 6,046,005; 6,087,095; 6,210,891; and WO 01/20039. Theextension products can be detected in any manner, such as byfluorescence methods (see, e.g., Chen & Kwok, Nucleic Acids Research 25:347-353 (1997) and Chen et al., Proc. Natl. Acad. Sci. USA 94/20:10756-10761 (1997)) and by mass spectrometric methods (e.g., MALDI-TOFmass spectrometry). Oligonucleotide extension methods using massspectrometry are described, for example, in U.S. Pat. Nos. 5,547,835;5,605,798; 5,691,141; 5,849,542; 5,869,242; 5,928,906; 6,043,031;6,194,144; and 6,258,538.

Microsequencing detection methods often incorporate an amplificationprocess that precedes the extension step. The amplification processtypically amplifies a region from a nucleic acid sample that comprisesthe polymorphic site. Amplification can be carried out by utilizing apair of oligonucleotide primers in a polymerase chain reaction (PCR), inwhich one oligonucleotide primer typically is complementary to a region3′ of the polymorphism and the other typically is complementary to aregion 5′ of the polymorphism. A PCR primer pair may be used in methodsdisclosed in U.S. Pat. Nos. 4,683,195; 4,683,202, 4,965,188; 5,656,493;5,998,143; 6,140,054; WO 01/27327; and WO 01/27329 for example. PCRprimer pairs may also be used in any commercially available machinesthat perform PCR, such as any of the GENEAMP® Systems available fromApplied Biosystems, for example.

A microarray can be utilized for determining whether a polymorphicvariant is present or absent in a nucleic acid sample. A microarray mayinclude any oligonucleotides described herein, and methods for makingand using oligonucleotide microarrays suitable for prognostic use aredisclosed in U.S. Pat. Nos. 5,492,806; 5,525,464; 5,589,330; 5,695,940;5,849,483; 6,018,041; 6,045,996; 6,136,541; 6,142,681; 6,156,501;6,197,506; 6,223,127; 6,225,625; 6,229,911; 6,239,273; WO 00/52625; WO01/25485; and WO 01/29259, for example. A microarray typically comprisesa solid support and the oligonucleotides may be linked to this solidsupport by covalent bonds or by non-covalent interactions. Theoligonucleotides may also be linked to the solid support directly or bya spacer molecule. A microarray may comprise one or moreoligonucleotides complementary to a polymorphic site within a nucleotidesequence

Examples

The Examples hereafter illustrate embodiments of the invention and arenot limiting.

Example 1 Restriction Endonuclease Enhanced Polymorphic SequenceDetection Using HpvCH4V And NlaIII

The effectiveness of restriction endonuclease enhanced polymorphicsequence detection was demonstrated using several restrictionendonucleases (REs), including HpyCH4V and NlaIII (purchased from NewEngland BioLabs, Inc). Both of these enzymes were separately tested inmultiplexed genotyping reactions for their ability to specificallycleave one allele of a given polymorphism while allowing PCRamplification of the remaining allele of the polymorphism. See Table 2for the polymorphisms tested with each enzyme.

Two CEPH DNA samples were mixed in varying ratios to generate DNAsamples composed of 0%, 2%, 5%, 20%, 50% and 100% DNA heterozygous forboth alleles of the SNP, with the remaining DNA being homozygous for theallele recognized by the RE. Table 3 shows DNA samples used in 5 thesestudies and corresponding genotype information. Mixtures composed ofNA05995 and NA10849 were used for experiments with HpyCH4V enzyme, andmixtures composed of NA10862 and NA10846 were used for experiments withNlaIII enzyme.

TABLE 2 Restriction enzymes recognizing SNPs Allele Restriction SNPDigested Enzyme Polymorphism Alleles by RE rs10430091 A/T NlaIIIrs2050927 A/T A NlaIII, HpyCH4V rs4329520 A/T T, T* rs4657868 A/THpyCH4V rs4658481 A/T A rs6693568 A/T rs860954 A/T rs9431593 A/T *Bothenzymes, NlaIII and HpyCH4V, digest the T allele.

TABLE 3 DNA samples used and genotypes Restriction SNP genotypes EnzymeDNA* rs2050927 rs4329520 rs4658481 HpyCH4V NA05995 TA TA NA10849 T ANlaIII NA10862 AT TA NA10846 A T *DNA samples were obtained from CoriellCEPH DNA collection

TABLE 4 DNA mixtures (Listed as ng DNA per reaction) Relative percentageunrecognized SNP allele 0% 2% 5% 20% 50% 100% HpyCH4V NA05995 0 0.6 0.60.6 0.6 0.6 NA10849 0.6 29.4 11.4 2.4 0.6 0 NlaIII NA10862 0 0.6 0.6 0.60.6 0.6 NA10846 0.6 29.4 11.4 2.4 0.6 0 NOTE: Based on 3 pg DNA forhaploid human genomic equivalent, 0.6 ng DNA is equal to 200 copies ofgenomic target DNA in the mixtures.

After preparation of the sample DNA mixtures, PCR cocktail was preparedaccording to Table 5 below (using multiplexed PCR primers as shown inTable 6) to either include no restriction endonuclease or 0.25 U ofrestriction endonuclease per each sample reaction. PCR cocktail wasaliquoted to a 96-well plate to include 7 replicates of each DNA mixturefor each enzyme condition. After addition of DNA to the PCR cocktailmixtures, samples were incubated at 37° C. for 1 hour to allow enzymedigestion of DNA samples and then immediately thermal cycled usingstandard conditions (Table 7).

TABLE 5 PCR cocktail preparation for each multiplex without DNA additionNo RE HpyCH4V NlaIII N = 1 N = 1 N = 1 Reagents Final Conc (uL) (uL)(uL) Water n/a 3 2.95 2.975 10x PCR Buffer 1.25x 3.125 3.125 3.125(HotStar Taq Buffer) MgCl₂ (25 mM) 1.625 mM 1.625 1.625 1.625 PCRNucleotide Mix 0.2 mM 0.5 0.5 0.5 (for UNG use) (10 mM dATP, dCTP, dGTP,dUTP) F/R Primer mix (0.5uM) 0.1 μM 5 5 5 5U/ul HpyCH4V or 0.25U/rxn —0.05 0.025 10U/ul NlaIII 1 U/μl Uracil-DNA- 1.25U/rxn 1.25 1.25 1.25Glycosylase (UDG) HotStar Taq (5U/uL) 2.5U/rxn 0.5 0.5 0.5 DNA - addedseparately varies 10 10 10 Total volume n/a 25 25 25

TABLE 6A PCR Primer sequences for SNPs SNP Forward PCR Primer ReversePCR Primer rs10430091 ACGTTGGATGCACAAGATTCTGAAACTTAGACGTTGGATGGCTGTTTAACTCAGCATG rs2050927 ACGTTGGATGTTGGGTGCAGAGTAGTCATCACGTTGGATGTTCTAGCTTGCTTCTCCTCC rs4329520 ACGTTGGATGATGTCCACCTCCTGCTCCACACGTTGGATGGAAAGTTGTCGTGGTAGAGG rs4657868 ACGTTGGATGCTAGCGTACCCAATGGAATCACGTTGGATGCTAACCAGGAAAAGACACCC rs4658481 ACGTTGGATGGTGGTAGAAACAAATGTCAGCACGTTGGATGCTGCTAAGCATGAGAGAAAG rs6693568 ACGTTGGATGGGCCTGTTCATTCTCAGAAAACGTTGGATGTGACTAGGAAATCACACTGG rs860954 ACGTTGGATGTAGCCTTTAGTCTTGATGCCACGTTGGATGCCATTCTTGTATGTTTTGTC rs9431593 ACGTTGGATGGCCTCAGTAGTCACATAAGGACGTTGGATGTTGAGATCAGTGTCGGTTCC

TABLE 6B Extend Primers SNP Extend Primer rs10430091gTGTTTAACTCAGCATGTGGGAA rs2050927 CCTCCATCATCCTTAGC rs4329520GCGTGGTTCTAGACTTATGC rs4657868 cAAGACACCCCCATACATTA rs4658481TAAGCATGAGAGAAAGGGAAAG rs6693568 atGAAATCACACTGGACATTTT rs860954GTTTTGTCTTTTTCTGTATACTCATG rs9431593 TGTTCCTGACTCTCAAAAT

TABLE 7 Thermal cycling conditions Temp. Time Cycles 37° C.  1 hour  194° C. 15 min  1 94° C. 20 sec 56° C. 30 sec {close oversize bracket} 45cycles 72° C.  1 min 72° C.  3 min  1  4° C. forever  1

Amplicons generated during PCR were genotyped with the extend primers inTable 5 using standard iPLEX™ assay and MassARRAY® technology (Jurinke,C., Oeth, P., van den Boom, D., MALDI-TOF mass spectrometry: a versatiletool for high-performance DNA analysis. Mol. Biotechnol. 26, 147-164(2004); and Oeth, P. et al., iPLEX™ Assay: Increased Plexing Efficiencyand Flexibility for MassARRAY® System through single base primerextension with mass-modified Terminators. SEQUENOM Application Note(2005), both of which are hereby incorporated by reference).

Results

Digestion of DNA with both restriction enzymes allowed detection ofminor alleles when they were present at ratios as low as 2% heterozygousDNA. This is in contrast to undigested DNA samples where minor alleleswere only reliably detected when present at ratios of 20% heterozygousDNA and higher. When allele peak area ratios are considered, the effectof restriction endonuclease digest is even more apparent. HpyCH4Vdigested samples showed minor allele peak area ratios of 0.35-0.45 in 2%heterozygous DNA mixtures, while minor allele peak area ratios of 2%heterozygous DNA mixtures were at background levels without enzymedigestion (FIG. 1). While the increases in allele peak area ratio werenot as high when using the NlaIII restriction endonuclease, the resultswere similar (FIG. 2). Example screen shots of the mass spectrum in 2%heterozygous DNA mixtures with and without HpyCH4V (FIG. 3) orNlaIII(FIG. 4) are shown below.

Optimization Studies

Initial optimization studies for enzyme concentration and pre-PCRincubation time of HpyCH4V digestion were performed using 5%heterozygous DNA mixtures (0.6 ng heterozygous DNA, 11.4 ng homozygousDNA). Based on these experiments, maximal peak area ratios were obtainedwith incubation times as short as 5 minutes and 0.25U HpyCH4V enzyme.

Example 2 Restriction Endonuclease Enhanced Polymorphic SequenceDetection Using Tfii

A similar experiment was performed as described in Example 1 using adifferent restriction endonuclease, TfiI. In this experiment, the TfiIrestriction endonuclease selectively recognized and cleaved the ‘C’allele of the ‘C/T’ SNP, rs4487973. The SNP rs4487973 occurs in thefollowing genomic sequence on chromosome 1:CACACAGTTAGGATT[C/T]ACCTGAGCTTGTCCC. For these studies, two CEPH DNAsamples, one homozygous ‘C’ and the other heterozygous ‘C/T’ for thers4487973 SNP, were mixed in varying ratios to generate DNA mixturescontaining 0%, 1%, 2.5%, 10%, 50% of the rs4487973 ‘T’ allele. The TfiIrestriction endonuclease was either added or not added to each mixtureto determine the endonucleases' effect on detecting the polymorphicsequence. Of the mixtures not digested with TfiI enzyme, the rs4487973‘T’ allele was detected in the 10%, and 50% ‘T’ allele mixtures, but notthe 0%, 1%, and 5% ‘T’ allele DNA mixtures. However, of samples digestedwith TfiI enzyme, the rs4487973 ‘T’ allele was detectable in 1%, 5%, 10%and 50% ‘T’ allele mixtures. These results indicate the utility of thismethod to improve detection of polymorphic alleles present at lowrelative concentrations in a sample.

Example 3 Fetal Identifiers Sex Test And Copy Number Determination

Selection of SNPs

Analysis of paternally-inherited alleles in clinical samples andcorrelation with Y-chromosome frequency in male fetuses was performedwith a total of 16 SNPs. SNP assays for analysis of clinical sampleswere multiplexed as 8-plexes. All SNPs had a minor allele frequency(maf) of ˜0.4 in all ethnic groups and were unlinked.

For performance evaluation of a universal Fetal Identifier panel thatcan be multiplexed with disease-specific markers, a new panel of 87 A/TSNPs with a pan-ethnic maf >0.4 was selected and multiplexed into16-plexes.

Method of SNP Analysis

Analysis of SNPs in maternal buffy coat and maternal plasma wasperformed using the iPLEX™ assay and MassARRAY® technology. In brief,the target region surrounding the SNP is first amplified by PCR.Subsequently an oligonucleotide primer is annealed to the PCR productand is extended allele-specifically by a single nucleotide using amixture of 4 terminator nucleotides and a DNA polymerase. The extensionproducts are transferred to a miniaturized chip array and are analyzedby MALDI-TOF Mass Spectrometry. Determination of the molecular mass ofextension products allows unambiguous identification of the SNP allelepresent in the sample. The peak area ratio of mass signals allows theestimation of the relative abundance of the alleles in a given sample.FIG. 5A provides an overview of the assay used for SNP analysis.

Clinical Samples

The total sample set consisted of 35 paired blood/plasma samples frompregnant Caucasian woman (nine 1st trimester; twelve 2nd trimester;fourteen 3rd trimester). The subset of samples used for correlation ofY-chromosome frequency and paternally-inherited alleles in maternalplasma consisted of 19 samples of pregnant Caucasian woman carrying amale fetus.

DNA Extraction

DNA extraction was performed from 1 ml of maternal plasma using theQiagen® MinElute kit for fetal genotyping. DNA extraction from frozenblood (minus plasma) was performed from 4 ml using Qiagen's PureGene kitfor maternal genotyping.

Results

An assay targeting sequence differences in the Amelogenin region on theX and Y chromosome was used to assess the relative amount of fetal DNAextracted from plasma of pregnant woman carrying a male fetus. Detailsof the AMG assay are depicted in FIGS. 8A-8C. X and Y-specific sequencescan be discriminated by sequence specific iPLEX extension products andtheir respective mass signals. The peak area ratio of the extensionproducts allows estimation of the relative amount of fetal DNA, becausethe Y-specific sequences represent 50% of the total fetal DNAcontribution.

Sixteen of nineteen (84%) plasma samples with a male fetus showed aY-chromosome frequency of higher than 5%, indicating presence of atleast 10% fetal DNA in the extracted DNA. FIG. 6 depicts typicalperformance results for a qualified fetal identifier. Here the abilityof the SNP assay to estimate the quantity of fetal DNA in the backgroundof maternal DNA was verified for a total of 1700 copies and a total of170 copies using genomic DNA mixtures. Note that the standard deviationof the estimate of fetal DNA increases due to the significant influenceof the sampling error at low copy numbers

Table 8 provides a list of SNPs that were multiplexed at 10+ plexinglevel and passed all phases of the validation. The following shows thevalidation scheme, performance criteria and model system used to qualifymultiplex SNP assays for their utility in identifying the presence forfetal DNA:

Phase I

-   -   Step 1: Initial Fetal Identifier (FI) screening parameters        -   FI's are multiplexed from pool of 87 A/T SNPs (mass            difference 56 Da)        -   Genotyping of control DNAs (CEPH populations)    -   Step 2: Advance screening criteria        -   Reproducibility of genotyping calls in 4 replicates        -   Unambiguous genotype data (assay shows no interfering or            unpredicted mass signals)        -   Allelic skew in heterozygous DNAs        -   Variance of allelic ratio in heterozygous DNAs    -   Step 3: Replex successful SNPs and repeat Phase 1 screening to        generate multiplexes of 10+ SNPs

Multiplexed SNPs passing Phase I test criteria are tested in Phase II

Phase II

-   -   Step 1: Mixtures of Genomic DNA are used for assessing FI        reliability        -   Mix Mother: 2000 copies of DNA1        -   Mix 10%: 3600 copies DNA 1/400 copies of DNA 2        -   Mix 20%: 1600 copies DNA 1/400 copies of DNA 2    -   Analysis of allele frequency variation in 4 mixture series and 8        replicate measurements. Sensitivity and specificity are        calculated for the detection of low copy number allele in        background of high copy number allele

Multiplexed SNPs passing Phase II test criteria are tested in Phase III

Phase III

-   -   Step 1: Various DNAs are mixed to emulate different        maternal-fetal combinations        -   Plate 1: 3600 copies DNA maternal/400 copies DNA fetal        -   Plate 2: 1600 copies DNA maternal/400 copies DNA fetal        -   Each plate contains 88 sample mixtures, 4 positive and 4            negative controls. Analysis of allele frequency variation in            4 mixture series, where sensitivity and specificity are            calculated for the detection of low copy number allele in            background of high copy number allele

Application of this assay panel to a model system for the detection offetal DNA in maternal background showed that paternally-inherited fetalalleles can be detected with a sensitivity of 95% at 100% specificity ifthe sample preparation method can enrich the relative amount of fetalDNA to 20%. In Table 8, the minor allele frequency (MAF) for each SNPfrom different ethnic populations is provided. The ethnic populationsare defined by the HapMap Project, where CEU represents individuals ofNorthern and Western Europe descent, HCB represents Han Chinese inBeijing, JAP represents Japanese in Tokyo, and YRI represents the Yorubain Ibadan, Nigeria.

TABLE 8 MAF MAF MAF MAF SNP CEU HCB JAP YRI rs11166512 0.43 0.41 0.500.49 rs11184494 0.50 0.40 0.48 0.50 rs11247894 0.43 0.39 0.32 0.44rs12089156 0.46 0.49 0.44 0.43 rs12125888 0.40 0.43 0.48 0.43 rs121363700.42 0.48 0.42 0.48 rs12143315 0.40 0.42 0.42 0.42 rs12759642 0.39 0.480.48 0.42 rs156988 0.46 0.40 0.45 0.41 rs2050927 0.44 0.50 0.41 0.49rs213624 0.48 0.44 0.40 0.34 rs2454175 0.46 0.48 0.43 0.40 rs43295200.45 0.43 0.40 0.44 rs4487973 0.47 0.43 0.44 0.40 rs454782 0.48 0.400.41 0.46 rs4648888 0.33 0.30 0.33 0.46 rs635364 0.49 0.40 0.46 0.43rs660279 0.41 0.49 0.50 0.39 rs6687785 0.48 0.46 0.48 0.44 rs75511880.46 0.49 0.45 0.46 rs9431593 0.41 0.43 0.49 0.40

A multiplexed panel of 16 SNPs was analyzed with maf>0.3 in the samematernal plasma DNA extraction and established a baseline of maternalgenotypes by analyzing DNA from PBMCs. Using the maternal genotypeinformation, paternally-inherited alleles were identified in plasmasamples and estimated the amount of fetal DNA from the peak area ratioof extension products representing paternally-inherited fetal allelesand maternal alleles.

The AMG XY frequency was then compared with the allele-frequency ofpaternally-inherited fetal alleles in informative SNPs. This comparisonrevealed that samples with a positive Y-frequency of 10% (used as aLimit-of-quantitation threshold) or more have significantly higherdifferences between maternally and paternally-inherited fetalallele-frequencies (p-value<0.001; Fishers' exact test). This datasuggests that Fetal Identifiers can be used as a non-gender specificapproach for identification of the presence of fetal DNA. FIG. 7exemplifies those results.

Example 4 Restriction Endonuclease Enhanced Polymorphic SequenceDetection Using Tsp509I

The effectiveness of restriction endonuclease enhanced polymorphicsequence detection was demonstrated using Tsp509I (purchased from NewEngland BioLabs, Inc). Tsp509I was tested in multiplexed genotypingreactions for its ability to specifically cleave one allele of a givenpolymorphism while allowing PCR amplification of the remaining allele ofthe polymorphism. See Table 9 for Tsp509I enzyme characteristics.

TABLE 9 Enzyme source E. coli expressing cloned Tsp509I gene fromThermus species ITI346 Recognition sequence 5′ . . . ↓AATT . . . 3′Vendor New England Biolabs, Inc. Catalogue Numbers R0576S, R0576L Stockconcentration 10U/ul Digestion temperature 65° C. Thermostable? YesTimesaver Enzyme? Yes Heat Inactivated at ≦80° C. No

Potential SNPs for use with Tsp509I

SNPs meeting the allele frequency criteria above were further screenedfor three characteristics:

-   -   1) one allele of the SNP is recognized by Tsp509I    -   2) the alternate SNP allele is not recognized by the Tsp509I    -   3) no other sites for Tsp509I are found ±50 bp of the SNP within        the PCR amplicon

338 SNPs passing these criteria are shown in Table 10.

TABLE 10 SNPs meeting criteria for Tsp509I screening rs10021843rs11221268 rs1447660 rs2367059 rs4130306 rs623052 rs7703746 rs10030074rs11221881 rs1458207 rs2373814 rs4311632 rs6431221 rs7725509 rs1003016rs11227624 rs1462685 rs2401505 rs4399565 rs644818 rs7737946 rs10034384rs11249671 rs1470207 rs2427102 rs4420242 rs6468296 rs7741525 rs1004395rs11563997 rs1503660 rs2435556 rs4420719 rs6488494 rs7763815 rs10102733rs11635372 rs1514424 rs2451984 rs4438888 rs6494229 rs7769867 rs1010479rs11655850 rs1536069 rs2462049 rs4442368 rs650616 rs7810506 rs10110766rs11685586 rs1540885 rs247852 rs4452041 rs6542638 rs7818415 rs10139699rs11727770 rs1543513 rs2507947 rs4488809 rs6556642 rs7820949 rs10179379rs11759755 rs1548605 rs2517540 rs4489023 rs6569474 rs7828293 rs10234234rs11771935 rs1593443 rs2522215 rs4533845 rs6575809 rs7831906 rs10260483rs11773909 rs1597205 rs263025 rs453609 rs6582294 rs7845628 rs1026791rs11835780 rs163027 rs264039 rs4589569 rs6592545 rs7899028 rs10276221rs12007 rs166576 rs2647415 rs4667489 rs6595267 rs7900002 rs10278812rs12034424 rs16830436 rs2657300 rs4673821 rs664358 rs7915178 rs1029176rs12107918 rs17074340 rs2676403 rs4674824 rs6707911 rs7985274 rs1041409rs12158945 rs17079191 rs269882 rs4678766 rs6766358 rs8016543 rs10421748rs12439908 rs17152417 rs2723307 rs4680921 rs6807437 rs8063107 rs10510379rs12442455 rs17156383 rs273172 rs4683161 rs683262 rs880385 rs1054067rs12450474 rs17170027 rs2734574 rs4684986 rs686851 rs910500 rs1070036rs1259733 rs1720839 rs2792780 rs4708590 rs6878291 rs9285190 rs10740169rs12607335 rs1789529 rs2804649 rs4716945 rs6897414 rs9312864 rs10754776rs12618834 rs179596 rs2820107 rs474077 rs691 rs9314663 rs10777944rs12674093 rs1797700 rs2821312 rs4762447 rs6929257 rs9322744 rs10784847rs12675087 rs1822243 rs2826737 rs4764597 rs6941784 rs9352730 rs10785736rs12783667 rs1850422 rs2828793 rs4783152 rs6962207 rs9356029 rs10795112rs12903747 rs1870836 rs2834712 rs4815732 rs7002630 rs9428474 rs10806232rs1297215 rs1885121 rs2846589 rs4845519 rs7041138 rs9515625 rs10818726rs13110085 rs1904161 rs2865878 rs4869315 rs7076662 rs9554894 rs10822434rs13130326 rs1904185 rs2889515 rs4889072 rs7082218 rs9555581 rs10832561rs13155942 rs1910369 rs2903113 rs4894467 rs7084321 rs9594249 rs10840805rs13255815 rs1912619 rs2928668 rs4897019 rs7094883 rs9599645 rs10851704rs13269702 rs1916803 rs2937415 rs4928169 rs7144509 rs9630712 rs10860857rs13331222 rs2007475 rs2984523 rs494220 rs7151741 rs9652080 rs10880400rs1335075 rs2030926 rs299080 rs4952502 rs7205009 rs9692857 rs10884498rs1342995 rs2034877 rs2993531 rs4953843 rs725849 rs9787011 rs10893402rs1346718 rs2038710 rs3010003 rs4974594 rs726395 rs9818611 rs10898954rs1363267 rs2063506 rs302137 rs514714 rs7266163 rs9838013 rs10901705rs1367452 rs2092797 rs309564 rs550408 rs7294836 rs9864594 rs10953770rs1372688 rs2126316 rs3128688 rs558692 rs7320201 rs9886292 rs10956363rs1376827 rs2168524 rs313937 rs561470 rs7323716 rs9929404 rs10964719rs1378933 rs2191076 rs331893 rs586030 rs7356482 rs9987005 rs10996924rs1401454 rs2207800 rs356643 rs6005754 rs748773 rs9989393 rs11017936rs1418136 rs2241491 rs373321 rs6019378 rs7588807 rs9992168 rs11079666rs1420562 rs2247858 rs3816551 rs6043856 rs7604667 rs11082446 rs1432865rs2298810 rs3902451 rs6139756 rs7679285 rs11099210 rs1439047 rs2304748rs3902595 rs614004 rs7688917 rs11105611 rs1444647 rs230526 rs3912319rs6142841 rs7689368 rs11125229 rs1445496 rs2322301 rs3913810 rs614290rs7691446

Multiplexing Tsp509I SNPs

Multiplexed assays were designed using 274 SNPs from Table 10. Theresulting multiplexed SNPs are shown in Table 11A with associated PCRprimers and Extend primer for each SNP, and genomic sequence comprisingthe amplicon sequence (with the SNP allele variants indicated bybrackets) are shown in Table 11B.

TABLE 11A SNPs and PCR and Extend primer sequences used for multiplexassays with Tsp509I enzyme Mul- ti- plex SNP_ID 2nd-PCR Primer 1st-PCRPrimer Extend Primer W1 rs644818 ACGTTGGATGTGTCAGACTTGTCTGAAGGCACGTTGGATGCAGATAGTGCTTGAGAGGAG GAAGGCCCACAGAAA W1 rs11685586ACGTTGGATGCCAAAGTGAACTTGGGTCTC ACGTTGGATGGGGAGAAAGAAACAACCTGCCGGGGACTCCAGGAA W1 rs7094883 ACGTTGGATGAAGCCTGTGGACTGTTAACCACGTTGGATGGACATTAAGCCCAAAACAGG AACCTGCTGACTTCAA W1 rs10021843ACGTTGGATGGTGAACTTTTTTTGCAAGGG ACGTTGGATGAAAGCTGGCCAGGGATATAGTTGCAAGGGAGGAAAA W1 rs7588807 ACGTTGGATGCATCAGCAGTGTGTAAGAGGACGTTGGATGCTGGTGAGTAAGCATTGAAG CTTCACCAGCACTAAGA W1 rs1297215ACGTTGGATGTCCAAGGTGGTCTTTTGGAG ACGTTGGATGGTTGGTAAATGGTAGAGCCGtcTGGCTCTGGGTTCAA W1 rs4667489 ACGTTGGATGTTTGTTAGCAGCTATGCTGGACGTTGGATGGGAGTAGTCTTCACCTGTAG cGCTATGCTGGAGCAAA W1 rs7082218ACGTTGGATGGTCTCTTAAGCAACGAGCGG ACGTTGGATGAGAAGGGCAACCAACAACTGCGGGTGCAGTGGGTGCAA W1 rs2903113 ACGTTGGATGACACTGTTCGCATCTGCATCACGTTGGATGGTAGCTCAGGCAAGGAGATT ccacTCCCAAGCCACAAAT W1 rs10139699ACGTTGGATGTGTTTCTCAGGAGTTCCCAG ACGTTGGATGGCAGGAGAGGAGAAAAAGACccaGAGTTCCCAGCAGAAT W1 rs4452041 ACGTTGGATGTGTGTCCAGTGACCATAAGGACGTTGGATGGTTGACGCAAAGCAAGTGAC tcCCTGTCAGTGAGGAAAA W1 rs13130326ACGTTGGATGTGGTTCCAGTTCTCAAGCTC ACGTTGGATGTCTTAGGAAACCACGTCCACCCTTTGATGAGGAGCTGTA W1 rs2865878 ACGTTGGATGATTGTGGCTGTGCTGTCCTCACGTTGGATGCGTATCTGTCTTGGATCCTG gagaGGGGACGATGCAGAA W1 rs11079666ACGTTGGATGGGAACACCTCCATTCTGATG ACGTTGGATGACACAAGTGGGAGAGGTTTGcctcGGGTCCTGGAACCCTA W1 rs2401505 ACGTTGGATGACACCATCTCGGTAGGAAAGACGTTGGATGCAGTTTGTTAGGTTCTCTGG AGTTTGACAGGAAGAAGAAA W1 rs4889072ACGTTGGATGCAGGAAGTATATGAGATCTGG ACGTTGGATGTACACAGTAAGTTCCCTGAGAGATCTGGAGGATGGAGAAA W1 rs7151741 ACGTTGGATGAAGGGTGAGGTGAGATAACGACGTTGGATGCTGGTTCCAGCACAAGTTTC AGATAACGTGATCCATTTAAT W1 rs10034384ACGTTGGATGGAGTGAGTCCTTTGATCCAG ACGTTGGATGCTACTTCCAAAGATTGTTGcGAAAGTGCATAGCTTGTTAA W1 rs1822243 ACGTTGGATGCTCACAGTGAAAGTGAACAGACGTTGGATGCCCGTATATGTAGCCACTTT tctcATGCTTTCAGCTCCAAAA W1 rs7604667ACGTTGGATGGACATATAATACCTTGGTCCC ACGTTGGATGCGTCTGCTTCCTTCATAGAGcaCTTGGTCCCTTATTGTTCAA W1 rs7845628 ACGTTGGATGGAGAGGTTGGGAAAAATGTGACGTTGGATGGGAAGATGCACCACTTTCTG gaaacTGTGAAGAAAGAGGAGG W1 rs7915178ACGTTGGATGAGCTTTCCTAAACCTGTGAC ACGTTGGATGAAACCACTTCCTGCTTTCCGctacACCTGTGACATTGGTTTAA W1 rs7691446 ACGTTGGATGACCACCATCACAAAAAGAGGACGTTGGATGTATGTTTGCATGTTGTTTG aaCATCACAAAAAGAGGCTCTAA W1 rs4684986ACGTTGGATGCCATGTGAGGAGGCATGTTT ACGTTGGATGTTAATGCCAGACAAGCCTCCtttggGGAGGTACATGAGGGAAA W1 rs4894467 ACGTTGGATGGTATTGGGTTACATGATGACGTTGGATGAGAAGGTCCTGTTAGTAGGG ATGATGTAATAACTAAAATGCAAT W1 rs16830436ACGTTGGATGGCGTGCATGGACTTCACAAG ACGTTGGATGCCACTGGCCTTTTCAAAGTCACAAGAAGAAATGTCTAGATTTAA W1 rs7810506 ACGTTGGATGGAGCATCTTCAAATATCCCCACGTTGGATGAACAACCGTTTTCTCTTGGG ctataCCCTTTAGAATGACAT TCAA W1 rs11017936ACGTTGGATGAATCCATTTCAGACGCAGCC ACGTTGGATGAATGTCAGAGATCACAAGCCttacCTCATCAATGCAATCTG GAAA W1 rs17170027 ACGTTGGATGGGAACTGATGGAAGAAAAGCACGTTGGATGCCTTTTGTGAGCAAGATGCC cAGAATAGAATAGGAACTCAG AAAA W1 rs1378933ACGTTGGATGTGGGCACTGTAATACAAAGG ACGTTGGATGTCCACACATGGTATCACAACgggcgTTCAATGGAGAAGACA GAAT W1 rs4438888 ACGTTGGATGCTGTTGCCTAAAGTTCTCGCACGTTGGATGACATTACTTGAGACCCACAC CTCGCTATTGTTAGCATTAATA AGAT W1 rs2846589ACGTTGGATGGTGATATTGAGTCTCACCTG ACGTTGGATGCTCTTTCTCATTATCATTCGAAAGCAAAATGTGTATTTTTA CAAA W1 rs2298810 ACGTTGGATGTGGTCCAGTAGGAAAACAGGACGTTGGATGTTCACTGACTCATGGATGGG cccctAAAACAGTTCGTATTTCA GAAT W1 rs2034877ACGTTGGATGGCATTTTGGGAAATAATACC ACGTTGGATGGGGAAGTCAGGATGAAAGTGcttgTGGGAAATAATACCACATC CAAT W1 rs269882 ACGTTGGATGTACCTTCTATATCCAAGGACACGTTGGATGATCCTCCCTTTTGAAACTTG ggGGACATAAAACTTCAATGATA AGAA W1rs10102733 ACGTTGGATGTCAGAAGGAGAAGTACCAGC ACGTTGGATGGCTAGGATTACACGTGTGAGgggcCCAGCCTTGATGTGGGGAA AAAA W1 rs1259733 ACGTTGGATGCTGTCTGTGTGATCATCAGGACGTTGGATGTGACGCTAAAGACTGAGTGG gatggCTGTGTGATCATCAGGGA GAAT W1 rs9555581ACGTTGGATGCATTGAAACCTGGGATACAC ACGTTGGATGAAAGGCAATCTCGACCTCACtctcTGCTGAGGTATCATCTCTAA GAAT W1 rs10510379ACGTTGGATGTGCTCACACAAAGCCTGTTG ACGTTGGATGGAATAACTATGAGCTCATGGggtcgCTTCACACGGACATGCGTG ACAA W2 rs11835780ACGTTGGATGTGAATCCCATGAGCATGAGC ACGTTGGATGATTCCACACAGCATTGCCTCGAGCCCACTGCTACA W2 rs166576 ACGTTGGATGGCCTTATTAGCTCTCACTTGACGTTGGATGCATCTCATGAGAAAGGCATC ACATGGTCGCCAAAA W2 rs880385ACGTTGGATGGAAAGGCCACAAAGCTGTTG ACGTTGGATGCACATGCATGAGTATGGGACACTGGCTGGGAAAAA W2 rs4708590 ACGTTGGATGTGCAGAGCTGCGAGAAGAAGACGTTGGATGAAGAGAAGGGCTTTGCATCC aCACTGCACAGCCAAT W2 rs13110085ACGTTGGATGAGCAAGTGTTCCCTTTTTGG ACGTTGGATGCACGCGTAGGCTATGGTTTAGGGGCTGGTAGGAAAT W2 rs1797700 ACGTTGGATGAAGTGCTGGGATTACAGGAGACGTTGGATGGAGACAGGCAAAGATGCAAC TGGCCAGAACTAATCAA W2 rs1885121ACGTTGGATGGAGACGATTCTTCAGGAAAC ACGTTGGATGCCATGACTCTAGTGACCTTCAAGACAAAGGACACCAA W2 rs1904161 ACGTTGGATGTAAGCATCCATGGACCTACCACGTTGGATGCAGGTGGTAAATGTGCTCAG caGACCTACCACCCAAAT W2 rs10901705ACGTTGGATGTCTGAAGGTAGACCTGGATG ACGTTGGATGCTCAGGATATCATTACACACCaCTGAGAGCAACCACTAA W2 rs7820949 ACGTTGGATGCGAGTTGAAGATCCCATACGACGTTGGATGCTCGGTGAACTATAGGAATC CATACGAGTGGGAGAAAT W2 rs7144509ACGTTGGATGAAGCAACTGGCACTCCTAAG ACGTTGGATGGAGTGTTGTGATGCATGCCTGGCACTCCTAAGACCAAA W2 rs8016543 ACGTTGGATGTTATACAGGTTCCAGCCAGCACGTTGGATGCAGAGAGAAAAGGGAGTAGG ACCTGATACTGAAGCCAAA W2 rs1458207ACGTTGGATGTCTCAAATATCTAAGTGGG ACGTTGGATGGCAAAACTTCACCTCAATAAggTCTAAGTGGGAGTCCAA W2 rs13155942 ACGTTGGATGCGGTTTCTTTTGAGGACTGGACGTTGGATGGCTCAGTGTCTGACAAAAGC ctcTCTTTCTCCAGGGATGA W2 rs3912319ACGTTGGATGACTGGCCATGCAGATGTAAG ACGTTGGATGCACTGCCCATAGACTCTTTCgCCAACAGAGAAAGTAACAA W2 rs9929404 ACGTTGGATGGAGATGAGTAAGAGCAGGTGACGTTGGATGCTCATAAGACCCTGAACACC GAGCAGGTGAAATGTTTCTA W2 rs4974594ACGTTGGATGGAAAAATCCATCCTCTGAACC ACGTTGGATGCCATGGCTCGTGTTCTTAACcTCCTCTGAACCTTATCAAAA W2 rs4673821 ACGTTGGATGGTCACTGAACTCTGGAGTAGACGTTGGATGGCAGTTTTCAAAGGAAACCC agCAGATAGCCTCTTGTGAAT W2 rs10784847ACGTTGGATGTCCCCCTACTTGCTTGAAAG ACGTTGGATGTGAAAGAGTGAAGGGAGGACggggaTTGAAAGCAGGGCATA W2 rs1444647 ACGTTGGATGCTCCCATCTATGATTTCCAGACGTTGGATGATGCATATCTGGAGACACAC ccacATCATGCCTCTATTGACA W2 rs12007ACGTTGGATGAATGAGAGCTTGCTTACTTC ACGTTGGATGAGTGTCGTTCAGACACTAGCctAGCTTGCTTACTTCTAAAAA W2 rs6569474 ACGTTGGATGCATTGCAGTAACTGGAGGTCACGTTGGATGGGCACAGTAGTTCAGTTACC gATCATTGTATAGGTTCCCAGA W2 rs7076662ACGTTGGATGAACACCAAGGAAAGCGGATG ACGTTGGATGCTGCTTAGTAACTTCTGTCCAGCGGATGAAGCAATACATTAA W2 rs6043856 ACGTTGGATGTAATACCCTGAGCAAGGACGACGTTGGATGGTGCATTTAAAATCCATGTG cccatGACGTCACCCTGTAAAAA W2 rs6142841ACGTTGGATGGTCCATTTAACGGTGTGGAG ACGTTGGATGGGTTCATGAAATGTTAGTTCCcccccGTGTGGAGAAGTGCGAGT W2 rs748773 ACGTTGGATGCACCAGTGCAAACACACAACACGTTGGATGCCTGATTGTTTTGGAAGGAG gaagtAATGGAGAACCTGGTTAA W2 rs1363267ACGTTGGATGTGTGCAGCACTTTTCACAAG ACGTTGGATGCAGGGTCACATCACAGATTGcccCAAGTTGAAAACTTATTCCAA W2 rs2723307 ACGTTGGATGGGATCAAGAGGAAAAAATGGGACGTTGGATGTAGTTTCAATCTCTGTGCTG cATGGGAAACATGCCTCAATAAAT W2 rs4589569ACGTTGGATGTACATTCAGACGATAGTGCC ACGTTGGATGAGACCAAGTAACCCCAAACCggtaAGACGATAGTGCCAGAAAAT W2 rs6766358 ACGTTGGATGCACATGCTAGAGAAAGAGGGACGTTGGATGTATGTCCTTCCCTGATTTTC ccctcAATCATTCTATGAAGC CAAT W2 rs7689368ACGTTGGATGAGTTGCCATGTTTCCACAGG ACGTTGGATGGACTAATACTCAGGTTGAGGccCAGGATCCTCTAGATTGTG AAAA W2 rs7900002 ACGTTGGATGCTACGTGACCCAAAGTTCAGACGTTGGATGTCTCACTCCTGGTTACCTAC ggggcCCCAAAGTTCAGGATG GTAA W2 rs4489023ACGTTGGATGGGGCTCTTATTATTGTACTC ACGTTGGATGAACAAGCCCAAGTTCTCCAGcGGCTCTTATTATTGTACTCTA TAAA W2 rs10260483 ACGTTGGATGAGAAGGAGGTCATTCTAGGCACGTTGGATGACATGGACTCTAAAGCCACC gggcGGTCATTCTAGGCCATTA ATAA W2 rs4533845ACGTTGGATGGGCAGAACAAGGACAGATAG ACGTTGGATGAGTCTAGTAAAAGTTCTGCCatcGGTGGATGTTTCAGGGAAG TAAA W2 rs6556642 ACGTTGGATGGCCAGCTTGTCCATTAAAGGACGTTGGATGCTGGCTTATAAATAAAAGACC cACTTGAAAAATACTTTAGACTT TCTT W2rs12674093 ACGTTGGATGTTTCACAGGGTTAGGATGGG ACGTTGGATGCTAGCAAAGGCTGGATTCTGacatGGAGTTTCCTGTACTTTAA AAAA W2 rs7741525 ACGTTGGATGTGGAAGGCAGAGTGATATACACGTTGGATGGCTTTCTTCACTCAGAAGGG agagACTGAGACAGGCAGTAGCC TAAT W2 rs2462049ACGTTGGATGGGGAAGGTGTTTGTCTCATA ACGTTGGATGTGGTACAGTTTGAAAGGAGCggtcCTTTCTGCAGCTCATATTCT GCAA W2 rs11105611ACGTTGGATGAGAAGATATGTTGAGAGGGC ACGTTGGATGTATTCCCTTTCTGGCTGTGGcccctAGAGGGCAGATAAATAGTT AAAT W3 rs2191076ACGTTGGATGGTATGGTGCCTCCACAAAAG ACGTTGGATGCCTCTGGATATATGTCCAGTACTGTTTGACCCAGG W3 rs163027 ACGTTGGATGATGGTGGTGGCAATATTGGGACGTTGGATGGCCAAAAAGCAGGCTTCTTC TGGGAGGGGGAATAA W3 rs4420719ACGTTGGATGACCATTTATTGGCCCTGCTC ACGTTGGATGATGGCAACATCTGCTTTCCCGGCACCTTAGGTGATG W3 rs2038710 ACGTTGGATGAGAATGACAAACCCAAGGGCACGTTGGATGGGACCTGTGCAAAACTTTGG tAGGGCACGTAGTAGA W3 rs1850422ACGTTGGATGGTAGGTTAAGAGGGAAAGGG ACGTTGGATGACTTGCCTTGTTCTTGACTGAGGGAAAGGGTGAAAA W3 rs1447660 ACGTTGGATGAAAGTCAGCACAGTCACTGGACGTTGGATGTCTCGAACAAGCTAGAGGAC ttAAAGCAACCCCAGGA W3 rs11221268ACGTTGGATGTCGAACTCCTGACCTCAAAC ACGTTGGATGCCTGTAATCCCAGCACTTTGGACCTCAAACAATCCAAT W3 rs4845519 ACGTTGGATGGTGTTCATACTGTAGGCTTGACGTTGGATGTAAACCAACCCCCTTCTTGC cCTGTAGGCTTGAAGAGA W3 rs1514424ACGTTGGATGGTGTAATAGGCTTGTGAGAG ACGTTGGATGCTCTTTGGATTAAATGCCTGCGGCTTGTGAGAGGTAAAT W3 rs2092797 ACGTTGGATGTGCTTCATAACTCTGTCACGACGTTGGATGCAAAACAGTATCGTAACAG tTCTGTCACGTTTCAGTAA W3 rs3902595ACGTTGGATGCAAGTCTCCCTAGCTAAGTG ACGTTGGATGTAGGAAGATCCTGGAAGGTGgTCAGATCAACACCAAGTA W3 rs10276221 ACGTTGGATGCATTTGCGGCAAAGAGGGAGACGTTGGATGAGCTCCCACACATGAAAGAG GGAGCCAGAAGGATATAAT W3 rs11249671ACGTTGGATGCACCCTATGCGACTTCTTTG ACGTTGGATGGTGGAGCTGTTATTCTAGTGtttcCCACCGTCGAGACAAT W3 rs9992168 ACGTTGGATGTATCCCCCAAACCTCACATCACGTTGGATGGAGTGGACTATAGTGGATGC CCAGAGGATGTGTACACTAA W3 rs2937415ACGTTGGATGATCATGGAAGTGATGAGAGG ACGTTGGATGGCCACATTCAACTGCAGTTCGAAGTGATGAGAGGAACTAA W3 rs10421748 ACGTTGGATGAGGACCTGGAGCTCAGCAACACGTTGGATGCTCAGCTGTCTCCATGCTC ggggTGGGGAGAATGCCAAA W3 rs11227624ACGTTGGATGTGTGCAGCAATGATCACAG ACGTTGGATGCTCAGCCATCTCCTGTCATCAGCAATGATCACAGCTATAAT W3 rs6595267 ACGTTGGATGACAAGTAAGGTTGGGTGGTGACGTTGGATGCCTATTCATGGAACCTCCAC ggaaGTTGGGTGGTGCCTTTG W3 rs614290ACGTTGGATGGGATGCTATATCATAGCCAC ACGTTGGATGCTTCCCCCGCTCTTTTAAACCCACATACCTTGAAAAAAGAAT W3 rs1536069 ACGTTGGATGCTCTGCTCTGCACACATAAGACGTTGGATGCCCTGAGATTATGTGACACC gctaTGCACACATAAGGAGTAA W3 rs7688917ACGTTGGATGGGTGTTAGTCAACTAGGAGG ACGTTGGATGAGAGCTTGGACTCTAGCATCTAGGAGGTAATGGAGAAATAAT W3 rs4420242 ACGTTGGATGAGAGGAAGCAAAGCTAAGGGACGTTGGATGCCCAGACCACTTTATAAGCC ccctcCAGATCCAGAAACAGGAA W3 rs1432865ACGTTGGATGAGGAGGTGACATTTAAGCTG ACGTTGGATGCTTTGCACTTACTGCTTCCCggacACTGAATGACAAGAAGGAA W3 rs2821312 ACGTTGGATGACGGCTAATGCTCCTCATTCACGTTGGATGGCATGTTTAGTACCTGCAAG ctccCCTCATTCAACTCAATGTAA W3 rs1910369ACGTTGGATGGGGCTTGAATAGCTAGATAC ACGTTGGATGTTACCTAGCTAGAGATCTGGGCTTGAATAGCTAGATACCCAAAT W3 rs2030926 ACGTTGGATGGATAGGGATAGACACAGGACACGTTGGATGGTAGTTAAAGGTGAGCAGGG atagcCACAGGACAAGAAACCAAA W3 rs4399565ACGTTGGATGGGATTTCTGTGAAGCTGCTC ACGTTGGATGAAAGTGTTGACCCCAGTGTGtggaTGCTCTAGAGATGAGGACAA W3 rs1367452 ACGTTGGATGCCATGAATGGCAAGTGTCTGACGTTGGATGCTTGGGTTCTGAGGATTTGC cccccCTTCAGGCCAAATCGA GAAT W3 rs2828793ACGTTGGATGTTGGTAGCATATGGGTCTCC ACGTTGGATGCCTTTTCTGATGAATGAAGCCctGTAGCATATGGGTCTCCTT TTAA W3 rs2427102 ACGTTGGATGGCCAGGGATTGTATTCGAAGACGTTGGATGCTGGATATTGTTCAGCTGGG gggtTCAGGAAGCTCTGGAAT CAAT W3 rs10860857ACGTTGGATGCTTCTATGAACCACCAAGGC ACGTTGGATGGGATACAGCCAAACCATGTCccacaACCAAGGCAAGCGACAA AGTC W3 rs9692857 ACGTTGGATGGGTAGGAAACGTGTACACTGACGTTGGATGATCCATGAAAACAGGATGTC AAACTATAAAGCATTGCTAAAA GAAT W3 rs4762447ACGTTGGATGATGCCTATTTCTTGTGACCC ACGTTGGATGCTATACTGCACCTTAGAACCgggagGGTTTTTTGCCAGTATG TAAA W3 rs1540885 ACGTTGGATGATGACATACTCCCATGTGCCACGTTGGATGGAAGAAGAATCAGAGCCAGC gaagGTGCCCCCCAGGTTTTGAA CAAT W3rs17156383 ACGTTGGATGCACGCTATGTAAAAGTAGCA ACGTTGGATGCTTCCAAAGTTCATATGCAGccAGCTACTGAAAATGAAAATGT ATAA W3 rs10278812 ACGTTGGATGGAATGGATAGAAGAATCTGACGTTGGATGACTACCCTGACTGCTATCTC gggcATGGATAGAAGAATCTGTC ATAA W3 rs6575809ACGTTGGATGCCTGAGTCAACCTTGGAAAG ACGTTGGATGTAATAGCTCCCCCAACAGTCgGGAAAGATAAGAGAGATATCAG AAAT W3 rs1029176 ACGTTGGATGAGCCTGAATCTCTAGCAGTCACGTTGGATGGAGAGACACTGTCTCACTCA aaggCATAAATATGCTTTCAACTA CATG W3rs3010003 ACGTTGGATGCTGCAAGCTAAGAAACACAC ACGTTGGATGCGTACCATATACCTAGGGTGgcttGGTGATTTATGCAGAAAAAG AATA W4 rs2241491ACGTTGGATGCCATTATTTCTCCCAAAGCTC ACGTTGGATGAAATAAGACCCTTGCACCCGCCAAAGCTCTCCCAA W4 rs11635372 ACGTTGGATGTTGTGGAAGGAGGCAAGGGACGTTGGATGATGTCTGTCTTGGCTATGGG TCTGCAGTCTGGCAA W4 rs4952502ACGTTGGATGAGAAGAGATGGTGGTTGTGC ACGTTGGATGACTGTTAGCTAGCACTGTGGTGGTTGTGCAGCCAA W4 rs2063506 ACGTTGGATGAGTATCCTCCAGTTTAAGGGACGTTGGATGGGACTCCCTACTCATTCAAG GCCTCAGGGGAAGGAA W4 rs650616ACGTTGGATGGTTGTTGCTAGTAGACCGAG ACGTTGGATGCTAGTTTTCTCTTCCCCAGCgCCGAGGGGTGGGAAT W4 rs10179379 ACGTTGGATGTTTAGTGACACCTCCCATCCACGTTGGATGGGGTAGTAGGAAGTGGTTAG CCAATCTGTCCGGAAAT W4 rs2657300ACGTTGGATGGGCATGCAACATAGACTTGG ACGTTGGATGTTAGTGAGCATCAGAGGCAGccTCCTAGACCTGTGCAA W4 rs9886292 ACGTTGGATGAGGCTTTCAGGATCTGCTTCACGTTGGATGCTCAAGGGCCATAGAAACAC ggGCTTCCCTGGGAAGAA W4 rs247852ACGTTGGATGGTGGACACAGGACAGCATTG ACGTTGGATGTCATCGCATCATGCATCCTCaCCTGGAAAGGAAGGAAC W4 rs2517540 ACGTTGGATGATGTGTCAAGACCATCTGGGACGTTGGATGACGGAGCAAGACTCTGTCTC tccaCAGTGGCTCCCAAAC W4 rs1335075ACGTTGGATGAGTTATTCTCCCGAGAAGGC ACGTTGGATGGCTAGGCAGATTGTGCTGTGCCACAATAGGATCTGCAAT W4 rs9630712 ACGTTGGATGGACATGGTTGTGTTGTGAAGACGTTGGATGAAGCACCGCTGGTGATAATG GTGAAGTAAAAGCTGGAAT W4 rs4928169ACGTTGGATGACTATGGGTAGTACATGGG ACGTTGGATGGCATCATTTGAATATTCACACtGGGGTCAGGTAAGGAATA W4 rs11771935 ACGTTGGATGTGCAAGCCCACAGGACAAACACGTTGGATGTTCTTGTGGATTCCACTCCG gacAACAAGTACCAGCAGTA W4 rs13255815ACGTTGGATGGTTGGTAATAGCTACAGCCC ACGTTGGATGAGAAGAGCTGACTGTCAGCGccatCCCTGGTCCCCTGGAAT W4 rs9838013 ACGTTGGATGTTTTTGTCCCCAAACATCCCACGTTGGATGTTTAGTGAGGGTGCTGGAAG ccACTACCATTGAGGTTTCAC W4 rs9599645ACGTTGGATGAGACATCAGAGAGAAGGGAC ACGTTGGATGGTATTAAAGATGAGCCCACAGGGACATACAAATCAGACTAAT W4 rs453609 ACGTTGGATGTTGTTCCTGACTTCAAGGGCACGTTGGATGACCAGTTCCTACCCATGAAG aTTCATAATGAAGCAGGAAAT W4 rs8063107ACGTTGGATGAAGGTGCTGTGGCAAGTTAG ACGTTGGATGCTGCTGTGGGTATTCAGTTCgggaTGGAGGGTTTTTCACAA W4 rs9594249 ACGTTGGATGTGGAGAAGAAACTCAAAAGACGTTGGATGACAGGGTCTGTACATTGCAG cCTCAAAAGTTTAGAACCTGAA W4 rs7828293ACGTTGGATGCCAGGTCTCAACACTGATTG ACGTTGGATGGCCATTATGTGAAATCAGCGgtaaCAAGTAGAGGTGCTGAAT W4 rs7041138 ACGTTGGATGGAAATACTTCCCTCGGGCTCACGTTGGATGAACCGCAGGTAAGGATTCAG TTCAGGCTTTAAATACCTTCAAA W4 rs10818726ACGTTGGATGCTTCCCTGGCTTCATTTTCC ACGTTGGATGCGATCTCCATCAAAAGAGGCCTTCATTTTCCAGGGTTGTTAAT W4 rs1548605 ACGTTGGATGAGAGATTGAGCTTCAGTCCCACGTTGGATGTCAGTCTTGTGTAGATAGGG AGTCCCCTAGTGTAATAGGAAAT W4 rs6139756ACGTTGGATGCCACTTACAGAACAGAAGGG ACGTTGGATGTATACCCATCCCCCAATGACcccccCAGCAGGCTGCCTTGAAAT W4 rs2126316 ACGTTGGATGTGCTGCTGGATTCAGTTTGCACGTTGGATGGAACACTTTAGGCCAATATCC gggTGCTAGTATTTTGTTGACAAT W4 rs1420562ACGTTGGATGTTGATATGAGCCTCTGAGAC ACGTTGGATGAGCTGAAGTTCGTGAGATCCgagagGAGCCTCTGAGACTGAAAT W4 rs2647415 ACGTTGGATGGGACGTGAGCAAGAAAAGACACGTTGGATGTGCTACGATTCAGTAATGAG ggGAAAAGACACTATGATGGTAAT W4 rs12607335ACGTTGGATGGTGGTCTATTGAGGCAATGG ACGTTGGATGAGGTTCATTTATGTGGTAGCccccTCTGACAACAAAAGGAA ATAA W4 rs9322744 ACGTTGGATGGACCCATGTCTGTCATACTGACGTTGGATGTGGAGCACTTTTGATGTG AGCATCATTAAAGTATTTAGC CAAT W4 rs9864594ACGTTGGATGTGTCAAAACCCCATCTCTAC ACGTTGGATGGGGCTCAAGTGATTTTCCAGcttaAAAACCCCATCTCTACTA AAAA W4 rs4680921 ACGTTGGATGGCCAAGCAACACTATGGTATACGTTGGATGAAGACCAAGTGAACTGTGCC gCACCTTTTAGTCTAAGGAGAG AAAT W4 rs4716945ACGTTGGATGAATGCCATTTCCTCAGGAGC ACGTTGGATGGAAGCATCTAAGCACAGCTCgggcAGAATGAGGTGCTCTTTT CAAA W4 rs1543513 ACGTTGGATGGACTGGTAGAGTAAGTTCTGACGTTGGATGATTCCACATTCAGAGACAAC ggggTGTTTAAAGCAGGCAAAA TAAA W4 rs7266163ACGTTGGATGGTGTTGATCTGTCACATGGC ACGTTGGATGGAGAACAAATAGCCCTGAAGcttcCACATGGCAATATAAATGA CCAA W4 rs9515625 ACGTTGGATGGAGGTGCCAGCTAATCTAACACGTTGGATGCATGAGGCCACAAAGGAAAG cCCGTGATTTACTAATAAGTATC AAAT W4rs10953770 ACGTTGGATGTATTACATCGAAATCAAGG ACGTTGGATGAGGCAAAATCGTTTTCATCCgACATCGAAATCAAGGTTTATGT TATA W4 rs1070036 ACGTTGGATGGAAGTGTTTAGGATTTGAGACGTTGGATGTGCTCACTGGAGCATTTCAG cctcATACTTAGGTTGATTATCCC TAAT W4rs10822434 ACGTTGGATGAAGTCTTGACATAAGGTAG ACGTTGGATGGGCAATCTTAAAGAGGGTTGttacGTCTTGACATAAGGTAGTAT AAAT W4 rs4953843ACGTTGGATGCAAAAGCTTTGCGCATCAGG ACGTTGGATGACAGGACCCTTGCTTTCAACaggcgCAAAATCTAAAGCAGAGAT AAAT W5 rs2804649ACGTTGGATGTTAGGCCAAGCTCATGCTTC ACGTTGGATGAATCTGGCCAGGGAAGGTTGTCTTTCCAGGCCCAA W5 rs7323716 ACGTTGGATGCAGTGGATTTCAAATCCGGCACGTTGGATGTGTTCAGAGGGTGTTGGATG GCCGCACATCAGAAT W5 rs10785736ACGTTGGATGCAATCAGCTACTGCTGATCC ACGTTGGATGTGGTTTGGTTTCTCAGCTGGTGATCCACTGGCTCAA W5 rs7831906 ACGTTGGATGCTGTCAAAAGCCAGGCTAAGACGTTGGATGGAGGTTCAAAGAGTATAAAG CCAGGCTAAGGCAAAT W5 rs2928668ACGTTGGATGGCAACCAGTTATCCCCATTC ACGTTGGATGGTACTTTGTGACCTTGAGGCcTCCCCATTCCACAAAT W5 rs12903747 ACGTTGGATGGCTTGCAGAGGTTCACTAACACGTTGGATGTGAGGCCATTAAAAGCAGGG gATACAGCTTGGCCAAT W5 rs4869315ACGTTGGATGTAGAGCTCACAGAGCACTTC ACGTTGGATGAGCACTTAACTGAGTCTGGGGCACTTCCCTACAAACAA W5 rs6542638 ACGTTGGATGCTCAGTTTAAAGTCACTGCCACGTTGGATGTAACCCTGCAAAGACTAGAG cCACTGCCAGTGACCTAA W5 rs686851ACGTTGGATGTTTACAGACTAGCGTGACGG ACGTTGGATGATCTCACGATCCCCCATTTCcGACGGACCCAATCTAAT W5 rs9987005 ACGTTGGATGGGAGGATGAAATCAGTGGTCACGTTGGATGAGAACATGCCAGAAAGTGCC GGTGTTGCCTGTTATTGA W5 rs10030074ACGTTGGATGTTTTTCTGTCTCAGCCTCCC ACGTTGGATGATGGAGAAACCTGTCTCTACggaaACCAGGCCAGGCTAA W5 rs1346718 ACGTTGGATGTATGGATGCAAGCCTTTCCCACGTTGGATGAGGCTGAAGAATGCTTTCCC GACTATCCTCTTCAGACCAA W5 rs2007475ACGTTGGATGAGCTTGGGCTGAATGTTAGG ACGTTGGATGTAAAAGCAAAACAGCTTCCCctAGCGTTTCACGTTCAAAA W5 rs10110766 ACGTTGGATGGGCTCTAGTTTTCAGCAGACACGTTGGATGCTCAAAACCTGGCTACCTTG gCCTGGGAGAAAGAAAACAA W5 rs11099210ACGTTGGATGGTTACACTGACAATCAAGGG ACGTTGGATGACTCTCATGTACCCTCTCTGcGAGGAGGGCAGAGAAGAAT W5 rs4130306 ACGTTGGATGAACTGATGGCTCGTACTACCACGTTGGATGGCTCTTTTCCCTATGATGTG tGTACTACCCAGTGGAATAAA W5 rs1401454ACGTTGGATGGATAATATTGTGCTGCATGCT ACGTTGGATGACCTTGTTCTGTGTGTGTGGgggtTGCTGCATGCTGTAAAT W5 rs179596 ACGTTGGATGCTGGATCTTACCTCCATAGCACGTTGGATGACTAGAATCGTGCAGAGAAC agCTTACCTCCATAGCATCTAA W5 rs9787011ACGTTGGATGGAGCACTTATCACAGGTCAG ACGTTGGATGGAAGGTGGGATAAACAAGGGgtaatTGCCCCTTCAAGTGAAT W5 rs9989393 ACGTTGGATGACTGAAGCATAACGCCTCTGACGTTGGATGGGTGCCCAAACATGTTATGC gaCTCTGGGACTACTAAGAAGA W5 rs664358ACGTTGGATGATCTTCATGTCCCAAGGAGG ACGTTGGATGCCAAGTTTATGAAACGTAGggatGGAAAAGCTGAAAAGGAA W5 rs7737946 ACGTTGGATGTCACGTCAGACTACACTGAGACGTTGGATGGGATTATAGGCATGAGCCAC tcagcCTACACTGAGCTACCACA W5 rs1342995ACGTTGGATGCATTGCTTGGGTCTTCTCAG ACGTTGGATGGGGTTCTGGCAGATATATCCcccccCCTTCCATGGGACTCATTA W5 rs4311632 ACGTTGGATGGGTTTATTGGAAATGAAGTCACGTTGGATGGATCCTACTTACTTCCAGTC tcTTTAAAGTGCTACATCTATGAA W5 rs13269702ACGTTGGATGAAGAATGGAAAGTGATGAG ACGTTGGATGCTAGGCTTGTTCACTATTTGcGTGATGAGATTTCTATCATACAA W5 rs2993531 ACGTTGGATGCACTGAGAGATACAGGAAAGACGTTGGATGCTTGTTTCCCCAACATAAGG agcGAGATACAGGAAAGTGTAAAT W5 rs1372688ACGTTGGATGGCTTGTTAAATGTGTGTTCC ACGTTGGATGTCCCTCAGTTTAGTTTTGTCtttcAAATGTGTGTTCCATCA TCTA W5 rs1720839 ACGTTGGATGGATGATGAAAGCATAAGTCACGTTGGATGGAGATGTTGCAAAGATGCAAG ATGATGAAAGCATAAGTCTTT TAAT W5 rs6582294ACGTTGGATGAGTGAGACTTAACCGTGGAG ACGTTGGATGCACCCCCACATTAGCAAAAGaaatTGAACTGTAGCAAGAAA CAAA W5 rs10234234 ACGTTGGATGCTTCTTTTCCCTGCATCATCACGTTGGATGAGGGAAGTGTTGTAGCATGG catccGTTTTTCCCTCTTGACT GAAT W5 rs11221881ACGTTGGATGCTGCCTATTCTTCTACGGTC ACGTTGGATGCAGAAACATGCTTGTAGCAGgtcgTCTACGGTCTTTTTCTTA TCAA W5 rs494220 ACGTTGGATGCTTTGCTCACAAGAAAGTTGGACGTTGGATGCCCCCAAGGCAATGATTTTC TTGGAACTATCGTTCAAAAAGT ATTA W5 rs9428474ACGTTGGATGTGGAGGCCACTGGATTAAAG ACGTTGGATGAGACACAGCTAGCACTTTCCggTTAAAGGAGACAATGTATGT AAAT W5 rs7294836 ACGTTGGATGACTCCCTACCTATCTCTTTGACGTTGGATGTCCACAGCCACTGAATAGTC gtcgCTTTGAAAAGCCTTAACCA TTAA W5 rs614004ACGTTGGATGTGTTACAGCAGCTAGTGTTG ACGTTGGATGCCTCTAATAGCACCCAGTTCtcGCTAGTGTTGCACTAATAAAA AAAT W5 rs2304748 ACGTTGGATGCACCAGTCCCCTCAAATAACACGTTGGATGGCAGTTCTTAAAGACCTCGG acaAGTCCCCTCAAATAACCTATC AAAT W5rs2435556 ACGTTGGATGCCCTAGGATTTTCAGAATGG ACGTTGGATGGGCTGACTCATTTGTTAGGGcacTGGTTTCAACTTAAAATCGCC AAAT W5 rs550408ACGTTGGATGGTGCTTAGGAAATGTTTGTTG ACGTTGGATGCGTGAATACATGAGAAAGGCAATGAAAGAGATATAATCATCTTA AAAA W5 rs7818415ACGTTGGATGGAGGAGTTATAAGACCTAGAG ACGTTGGATGACCATATCACAGTTGTTGGGtgaagGAGAGCTTAACTAAAATAA ACAA W6 rs6488494ACGTTGGATGTATCCATCCTTCAGACACCC ACGTTGGATGATGGGACAGTAACTGCAGACAGACACCCAGGCCAA W6 rs10840805 ACGTTGGATGCCTACCTTGCTCTGAGAAACACGTTGGATGCTTCCTGCTTTTAAGCAGTC AGCCTGCACTGTGAA W6 rs11773909ACGTTGGATGTTTTTGGAAATGGCCCAAGG ACGTTGGATGGAAACAAGTAAATGAGGTCCTGGCCCAAGGAGAAAT W6 rs4764597 ACGTTGGATGAGATCCTCCAGCTCATCTTCACGTTGGATGTAATCCTTGGAGGCTCTCTG GCTCATCTTCCTCTGAA W6 rs2820107ACGTTGGATGAGATTGGTCCCTCACAATGG ACGTTGGATGATTTGGCCCTGAGGCTTATCAGTCTTTCTGAGCCCAA W6 rs13331222 ACGTTGGATGGGAATACATGTGGGTATGTGACGTTGGATGATATACGTTGCTTCCTTTGG GAGAGCCATGAGTGAAA W6 rs12675087ACGTTGGATGAGCCACCAAAACCAAGCTTC ACGTTGGATGCTTGTAAGGCAGGTCTGATGtgAGCAAGTGCTGAGGG W6 rs725849 ACGTTGGATGTTGTGTGCTATCTTACACTGACGTTGGATGACTAGTTGGAATGGGCTTGG cGTAGCTTCCTAGCCAAA W6 rs910500ACGTTGGATGACTGATACCCTACAGTGTGC ACGTTGGATGGTGCTCAGAGCACTTAAACGTGGATATGACTTGCCCAA W6 rs1916803 ACGTTGGATGTTGACTCACCCACTTCTGTCACGTTGGATGTGTTGATGAGGTGAAGAGGG ACTTCTGTCTCAGTATCCA W6 rs6431221ACGTTGGATGTTCAATCAGTCATGCCTGTG ACGTTGGATGCTAATCTGAAGGCTCCACTGcccTGCCTGTGTGATGAAA W6 rs4488809 ACGTTGGATGGCAAGCATCTGCTCTTGAGGACGTTGGATGCTGTGTAAAAGAGTTTGAGG cgGCTCTTGAGGCAGTAAA W6 rs3816551ACGTTGGATGGGTGGAGATGGGATTCTCTG ACGTTGGATGAACCCAGTCTACACACACAGGGGATTCTCTGGTTGTAAA W6 rs7205009 ACGTTGGATGGTATCTCCCACTCTTGTACCACGTTGGATGCTGGAATACAACATTTCTGG cCTCTTGTACCCCAGAAAAA W6 rs2322301ACGTTGGATGTTTTTCCTCCTGTACCCTGC ACGTTGGATGTACATGTGGTTAGAGTCTGGaaaTGCAATCTGTCTGGAAA W6 rs17074340 ACGTTGGATGAAATGCTACTCCAACAGAGGACGTTGGATGCTTCATTATCCCCACTGCTG GGAGGTGACATAAGTAAGTA W6 rs9356029ACGTTGGATGATCCTGGGCTTTCCTTTGTC ACGTTGGATGGAGTCTAGTGGACAAGAGAGacTTGTCACACCTCTTCAAAT W6 rs10898954 ACGTTGGATGAGTGCAACAGAAAAGGCAGGACGTTGGATGGGTCCTTGGTATGTGTTCTC CAAGTCTTCTATCAAGGGAAT W6 rs263025ACGTTGGATGGCATTATGCTAAAGGCTGTC ACGTTGGATGTCCTCTGATTTAGGCCCTTCGGCTGTCACAGATTTATAAAA W6 rs273172 ACGTTGGATGCTATGTTTTCCCCCAGCTTGACGTTGGATGGCAAAAGAACAACCACCCAG acttTGCTAGGTCTTACATGAA W6 rs9652080ACGTTGGATGGTTTGGTGACTATAGAAACAG ACGTTGGATGCAGTTTAAAGTCATATTCACgaagGTGTTGCCAAAAGCTAAT W6 rs2451984 ACGTTGGATGAACAATAGAGACACACTCCGACGTTGGATGTTTAATCCAGGGAGCTCTTC ccctcGACACACTCCGGCTAAAT W6 rs11655850ACGTTGGATGCACCACTCAGGAAAGCAAAC ACGTTGGATGAAAATCCCAGTGAAGAGCAGcccaAGGAAAGCAAACTGCTACA W6 rs7084321 ACGTTGGATGACAGAAGCACCACAGCTGAGACGTTGGATGAGGTTTCCCAAGCTAGACCC ttgacAAGACGCAGCTGTGCAAT W6 rs7356482ACGTTGGATGCATCAGCAATATAATGCCGC ACGTTGGATGTGTGGATCACTGTTCACAGGCAATCCTTTATCTCTCTCTAATAC W6 rs9818611 ACGTTGGATGGTTCTGGATGTTGGCCATTCACGTTGGATGCCACATCATATGCATCTGGG GTGCTATCTCATTGTTGTTTGAAA W6 rs7320201ACGTTGGATGTCATGTAACCAAGCACCACC ACGTTGGATGGCTCATTTATAGAAGCAGTCcccccCCCCAAAAACCTACTG AAAT W6 rs3913810 ACGTTGGATGTTACGACCCAATCACCTTGCACGTTGGATGTGTGTCCCCAACCACATTTC tCCTCCTCAAACATTAAGGAC AAAA W6 rs12450474ACGTTGGATGCCTTCTGCTCAACTACCAAG ACGTTGGATGGCCAAAGACGATGTGGAATGattaTGCTCAACTACCAAGTT AAGA W6 rs1503660 ACGTTGGATGCCAGTCAAGGAAGCAGTTTCACGTTGGATGGTCTGATTAGGCCTAAGAGC cCAGTTTCAATAACAGATAGT AAAT W6 rs683262ACGTTGGATGAGGATGCCTGTTGGGTTTTC ACGTTGGATGATCAGACTTTTCCCAGGCAGgTACTGAGATTGACAAGTCAT TAAA W6 rs1041409 ACGTTGGATGAAGCAGGTACTTACTATGGGACGTTGGATGGTACTGTTAGTGTGTCACTC cgggACTTACTATGGGGAATA GAAT W6 rs2984523ACGTTGGATGCTGAGGCACAAGGAGATAAG ACGTTGGATGACTGACCTGGGTTTGACTTCAAGGAGATAAGTAACATGTTTA AAAT W6 rs10754776 ACGTTGGATGTTGCCTAGCCTTACATCCTGACGTTGGATGCTCAAAATAGATGATGGACTG caccTCCTGAATACTTTCTCATA TAGA W6rs2734574 ACGTTGGATGGACCTTCCTGTTCCTAGATG ACGTTGGATGTGACTGGACTGTGACATAGCacgTTCCTGTTCCTAGATGATCA AAAT W6 rs9285190 ACGTTGGATGAATCTTGGAGCCTTGGAGACACGTTGGATGGTGCTTCTCACAAAAGCCTG gTAGTTTCTTTAGCTCTTGAATA AAAT W6 rs331893ACGTTGGATGATCCATCTCTGTCAGAGTTC ACGTTGGATGAGAGAACTGACCCTTCACTGaggTCTCAAATAAAAATGCAAAG GAAA W6 rs10806232ACGTTGGATGGAGAGAGGGAGAAAGTAGAG ACGTTGGATGCCCTTACTCAGTGATTCCTCggagGGTGGTTAGAGAACTCAAT GAAT W6 rs12107918ACGTTGGATGTTTAATAGGGAAAGTATTGG ACGTTGGATGCACACCCAGAAGCACTGATAaaatcTAAATAGCCAAGAAAACAG CCAA W6 rs1593443ACGTTGGATGTACCATGCTCATTGAACTCG ACGTTGGATGGGAGATTTGATAGGAAGTGCaaAAAACTCAATATAGTAAAGGTA TCAA W7 rs1912619ACGTTGGATGGCCCATCCTTCACTAACTTG ACGTTGGATGAACAGTGGTGGCCCATCAGTCACCCTCAGGAGGAA W7 rs1470207 ACGTTGGATGCTGCCAGGGAATAGGAGATGACGTTGGATGCGCTGAAAGAGACACTGAAG GGGTGGCATCGGAAA W7 rs7725509ACGTTGGATGTTACAGTTGAGAGCCACTGC ACGTTGGATGTGCCATTCATTGCTCTACACCACCGAGCTTGCAATA W7 rs2792780 ACGTTGGATGAAGTCATTTGAGGCCCATCCACGTTGGATGCACTTCCAGCTGCTGCTTTC gCCATCCTGGCTGAAA W7 rs6592545ACGTTGGATGTCTAGGTTGAGACTCAGGTG ACGTTGGATGGGTTTAAGCAACATGAAAGCcTGGCTGGACTGGGGA W7 rs6019378 ACGTTGGATGTACGACCAGAATGGAAGGAGACGTTGGATGATTGAACCCTGGGAAGGTGG cGAAGGAGGGCTTGGAA W7 rs313937ACGTTGGATGACTTAACCCCCAGTGTGATG ACGTTGGATGCACTTATCCCATTCACGAGGGTGATGGTGTTAGGGAA W7 rs10851704 ACGTTGGATGACTCTCACACAAAGTTTGCCACGTTGGATGCGGTATTGTCTTAAGACTGA CAAAGTTTGCCTGACAAA W7 rs6929257ACGTTGGATGGTCAGAGATTTCTGCCTAAG ACGTTGGATGCATCTGCCATGATGATCCTGTCTGCCTAAGGTGTTAAA W7 rs11082446 ACGTTGGATGACCCAGGTGACCGAATAAAGACGTTGGATGGTAGCTACGTTCTTTGGAGG TGGTATAGGTTTTGGGAA W7 rs4815732ACGTTGGATGGAGCTTCTATGAAACGTGTG ACGTTGGATGGGTTTCTGCCAAAAACCTTGcACGTGAAAACATGTTGAA W7 rs10795112 ACGTTGGATGCAGTCTATCTCTTGCTCTACACGTTGGATGGAAGACCATTATGTTTCTGAC TGCTCTACAATCACCTTAAT W7 rs1054067ACGTTGGATGCAGCAGCCTTTGAAAGACAC ACGTTGGATGTCAGCAGCTTACGGTTTCAGccTCAAACAAACAACAGACA W7 rs3902451 ACGTTGGATGCTGGTTCTGTGAAATAAGACACGTTGGATGTGGTGTCTTTACCTCTTTAC ATGAACAAAACCTTTGAGAA W7 rs9352730ACGTTGGATGGGTCACTAGTGTATATTTTG ACGTTGGATGGACTCCCAACACACAATACCAAGGGATAGTTGTAGTATGA W7 rs2207800 ACGTTGGATGCAAAAGAAGCTGGATTGCTCACGTTGGATGCCAAGAAAGGCAATGTTGGG ctCTGGATTGCTCTACTGGAA W7 rs1870836ACGTTGGATGGAGGAGAAGGTGATGTGAAG ACGTTGGATGCCCTGGAGTTCCTTTTCTTGATGTGAAGATGGAGGTAGAAA W7 rs264039 ACGTTGGATGATCCCTCATTCTTTCTCCACACGTTGGATGGAGAAGCTGAGGAAGCAAAG CATTCTTTCTCCACTAGATAAA W7 rs2826737ACGTTGGATGATGCTAAGGATTCTGGGGTC ACGTTGGATGCATATGCTAAGAGCCAGGACcttGGGTCAGACAGATTTGAAT W7 rs9554894 ACGTTGGATGGGGCATGACACAACTCAAACACGTTGGATGACCTGAGTTTTCAGCCGTTG ctgcCACAACTCAAACTTTGGAA W7 rs12034424ACGTTGGATGAACAGAGGGTTTAACAGCAC ACGTTGGATGACCTTACTCAGTTCTATTCTCAGATATGTTCAGTCAATGAAT W7 rs10880400 ACGTTGGATGCCAATATTTTTTCCCTAGGTACGTTGGATGTGTGCATTAAATCCTCCCCC ggggCCCTAGGTACAAAGGGCTA W7 rs9314663ACGTTGGATGTAAGCTCCCCCATCCAAGAC ACGTTGGATGCACAGGTCTACCTTGATTTCcccctCCATCCAAGACACTGGAAA W7 rs7769867 ACGTTGGATGTATCTGGGTCATTGTAAGGCACGTTGGATGTTCCCAAACATAATCACAG gCATTGTAAGGCAAATGTAATAAA W7 rs2522215ACGTTGGATGAGGGCACAAAGACATCAAAG ACGTTGGATGGGCATAGCGCCTGTGCTTAAgCATGCAAATCTTTCACATTA ATAA W7 rs10777944 ACGTTGGATGGGCAGGCACTCTATCAATACACGTTGGATGTGTTTGTTGCTGCGTGCTTC cagtCACTCTATCAATACAGG AATG W7 rs2373814ACGTTGGATGAGGGTATAGGAAACAGCTTC ACGTTGGATGATCCTCTCTCCTAACACCAGttcgTGTAAACAAGAGAAATC ATGG W7 rs6941784 ACGTTGGATGCATTTACCCACAAAGGTAAGACGTTGGATGTAGTCCCTGACATTGGAGAG GGAATGAAGAGATTAAAATAG ATAA W7 rs1597205ACGTTGGATGGATGCAGAATAAGCATTTGAC ACGTTGGATGGAGGCACTTTTTTCTGTTCCacTAAGCATTTGACAAAATCTG ATAT W7 rs11727770 ACGTTGGATGCTGTCTCAAGTGTCTGGTTCACGTTGGATGATCCATCCACCCATCCATTG ggCTGGTTCATAGTTAAAAGTC AATA W7 rs4678766ACGTTGGATGGTCCTAAGTTAAAAGAATGG ACGTTGGATGCTCATGCCGACAAAACTTCCCAAAGAAAAAGTAGATTTGTGA AAAA W7 rs3128688 ACGTTGGATGATCAAGAGGAAAATGGACAGACGTTGGATGGATTTACTCAACTCTCTGGG taatAGGAAAATGGACAGAAGT TGAA W7 rs2168524ACGTTGGATGTCTCCCCACTTTGTTCTGAG ACGTTGGATGTCAACTAAAGGGCAGTAACCTCAGCTACTCTGGTATTTAAAAT AAAT W7 rs11125229ACGTTGGATGACTGTGCCTGGACAAAGAAG ACGTTGGATGTAGCACCAGGCTTACTAGACgggaACAAAGAAGACCTGAAGTA CACA W7 rs6005754ACGTTGGATGGACAGTTTTTAAATCTTTTAC ACGTTGGATGCTGTATTCCCATACTACTTGTTTTTAAATCTTTTACATCAATAA CTAA W7 rs6962207ACGTTGGATGTGAGTGATAGGTCCTCTCTG ACGTTGGATGAGCTCACAAAACTAACACACGTCATTTTTTAAAATGGAAATCAA TAAA W7 rs12442455ACGTTGGATGCAAAAGAACCTGGCTCATGG ACGTTGGATGATATGTCACGCATAGCCCAGaaggcAGTCAGTGGATTATCCTTG GAAT

TABLE 11B SNP_ID Corresponding Genomic Sequence with Alleles provided inBrackets rs12007AATTTTAATCTTTGGTCTCTAAAAAGTAAATTTCAAATTTATGAGTTTAATCACTTCAAATATGAATAGCAAAAAATGAGAGCTTGCTTACTTCTAAAAA[T/C]TGAGGTTAAGATATAGCTAGTGTCTGAACGACACTCCTTAAAGTAAGTTCCAAATGTAAAACACTCCTTAAGTTCCAAATGTTTTCCGCTAATAGTCTGTrs691TACATGCATTCTTTTAGTGGATAGATGCACACAAACACACAAGCCATTATGGGGAAGGATCCACGTGTGTGGCCATATTGTAACACATTTTTCTGCAAAT[C/T]ACCTCTTTCATTTAACAGCCCTTATTCAATGGCCTTTTTCTTTTTCAGTAGTACATACACATCTGTGTCATTTGTTGAATGACGACATGAATGTTTTGTArs163027GTGAATCATGAAGTCATATCATCACTGTATTACAAAAGCCAAAAAGCAGGCTTCTTCATCTATCTTAGACTCACTGTGGTAGATCACAAACTGGCCACAA[A/G]TTATTCCCCCTCCCAATATTGCCACCACCATTAGCAATAGGACTTAGTTACTTCTACCATGAGAGGTCAAGTTTATTTACCCACTCACTGAATTTGTGCCrs179596AGAAGCACTAACCTAAACCAGTGGTTCTCAACTGGGCTGAATACTAGAATCGTGCAGAGAACTTTAAAAAATAGAGATGCCCAGGCTGAACCCCAAACCA[A/G]TTAGATGCTATGGAGGTAAGATCCAGACATTAGTCATTTTTAAGGCTCCCCAGGTTATTCTAACGCAAAAAAAAGAAAGTTGCCCTAAACCAGCTTTTTArs166576AAGTGATAATATTGCTATTACCTCCTGCATGCTGGATCTGTTTGCAGCCGTATTGCATGCCTTATTAGCTCTCACTTGTGTTTAAACATGGTCGCCAAAA[A/T]TCAGAGCCTTATGTTTGATGCCTTTCTCATGAGATGTAGGCCCACACATCCAACAGCCTGCTAGATATTGCCAATTGCATATCCTACACCTATATATGGTrs302137GATCAGTGATACTGAGCTTTTTTTCATATGCTTGTTGGCCACATGTGTGTCTTCTTTTGAAAAGTGTCTGTTCATGTCCTTTGCCCATGTTTTAATGGAA[C/T]TGACAAACCACAATCTTGATTGCTTAATACAATAAAGGATTATTTCTCACCATGTCACCATCCAGTTTGAGTTAGTATTCGTGGTAAGGGGACAGGTGTTrs264039TAGCCACAGCCATACAGGATAATTGCCCCATAATCTTTCACACCTCCAAGTTTTGGACAATCTAACAGAAAGATCCCTCATTCTTTCTCCACTAGATAAA[T/C]TCATTAATCCTTCAAGTCCCCTCTCACTTGTCATTTTTCTTTGCTTCCTCAGCTTCTCCATTAAGCTTTGTTTTTGCTTGTACATCCATTTCTTCCTACCrs247852CACCGGACTGGGAGCCACTGCGGGGCAGGAAGCTGCCTTTTCCATTTCCCAAGACCGGAATCAATCACAGCGGCTCATCGCATCATGCATCCTCTGCAAT[G/T]GTTCCTTCCTTTCCAGGGAGGTTGGTCACGGCCAATGCTGTCCTGTGTCCACCAGGTGGCGCTCGCGATCACCGCAAAACACATGGCTACCGTGATGGTTrs331893AATGATAAAATGTTTCGTTCTTTGGAAGTAACTCTTTTTTTTTCTTCTGTTCTTAGTCATCCATCTCTGTCAGAGTTCTCAAATAAAAATGCAAAGGAAA[G/T]TTAGCACCACTCTAAAACAGTGAAGGGTCAGTTCTCTGCTTTTGGATATGTAATTTGAATGGGAAGTGTCCTAATGACAATTAAACACAATTTTCTAAGCrs230526CTAAACTCTTCAGCAGATTACTCTCCACACATGCATAGCATGAGAGGTTCCATGGGCTTAGGTACCTGGCTTTTTAGCCATATCTTAGTGTACAAATATC[G/A]ATTAATACCATTTTTCGTAGTAAGATTACGGGAAAAGTGATTCTTGTTTACAGAGCCCTCTTTCAGTTTCATGTTTTTCTTCTCTCATTTAGTAGACATArs299080CTTTGAACCTGAGTGACTATTCCCACCTGATTCTTTTCTTTCTCTCCTTTCTTTTCCTCCCCAACATACGTTTATCGTCTCTTGCATTAGTGAATGGAAT[C/T;T/C]CGTATTCTTTCATGTAGAGAGCAACATCTTCCTACATAGTAAATAAAAGAGTAAAGACCACTGTATTGAGATGAGAAATCAAGGGAAGAAAGCAACCCAArs299080CTTTGAACCTGAGTGACTATTCCCACCTGATTCTTTTCTTTCTCTCCTTTCTTTTCCTCCCCAACATACGTTTATCGTCTCTTGCATTAGTGAATGGAAT[C/T;T/C]CGTATTCTTTCATGTAGAGAGCAACATCTTCCTACATAGTAAATAAAAGAGTAAAGACCACTGTATTGAGATGAGAAATCAAGGGAAGAAAGCAACCCAArs273172TCAATAATGTCTGAAGAGCTTGTAAAGAGATAAGTGAAAACCCTTGTAAGTATTTAGTTTCTTCTCAAACGCAAAAGAACAACCACCCAGGGCCCTTTTA[G/A]TTCATGTAAGACCTAGCACATGGAAATCTAGCAAGCTGGGGGAAAACATAGAGGCAGGAAATTCTGTGGCTTCTCTCAGAAGACAAGGATCGAATCCTTCrs263025GCCCTGAAAGGATGGTAGATAGGCCACATCTGCTTTTCTTTCGGAGGAAACAAATATTATTTTATTGCATTATGCTAAAGGCTGTCACAGATTTATAAAA[T/C]TGTAGCAAAGTGCCTGAGGACATGAAGGGCCTAAATCAGAGGAATAGTAGATGAAACTTCACAGGAATATATCAGACAACATAGGCGTCTTTAGAGAAAArs309564CTGAGTTTGACTCAGTCTCTGGAAATTTGCTGCATAAGTCTGGGCCCTGACCACCAATATCTGTCTCCTCATCCCTGCAATACTTTCTACTTGGACCCAA[C/T]TTCCACATGCTTGAATGTGGAAAAAAACGCTTTAGAGAAAAAGCAAGGGGAAATGTGGAGCTCACTTTGTATGTTTTGCTTCTCTCAAGGGTTACAGCACrs269882CTAGAAATAATGCCCTCTGGCCACTGCCCCAGTGCATTCTTATAGTGCACGCTGATAAATCATAGTAAAAATATATCCTCCCTTTTGAAACTTGCCTACA[G/A]TTCTTATCATTGAAGTTTTATGTCCTTGGATATAGAAGGTATTTTCATTTTCACCAACTCTGTAAAAAAAAAAACTACTTCTGGATTATATAAATACTATrs313937AGTTTAAGTAATATTTCAGAATGACACTGCTATGGCTTGAATATTTGTGTCCTCCAAAATCATGTTGAAACTTAACCCCCAGTGTGATGGTGTTAGGGAA[C/T]TGGGCCTTTAGGAGGTGATTAAGTCACGAGGGTAGAGCCCTCGTGAATGGGATAAGTGGCCTTATAAAAGGGGTGGAGGGAACTAGGTAGGCCCTTTTTGrs664358TTATAACACCTCCTTAACAGGTGTTTCACCTGATGCCCTGAGATGAAGTGCATTATCTTCATGTCCCAAGGAGGACATGGAGGGAAAAGCTGAAAAGGAA[C/T]TGTATTATGAAACTACGTTTCATAAACTTGGTAATACTACATTTTACAATATATGATATAACTACTGCTGATTAACCTGGAAAATGTTCATTTACTTTCCrs550408TTTTTAATGTGAAATGCTTGGCACAGTTCCTGGCTCAGAGTTAGTGCTTAGGAAATGTTTGTTGAATGAAGAAATGAAAGAGATATAATCATCTTAAAAA[T/C]TGTATTGAGTACTTATTATGAAGCCTTTCTCATGTATTCACGTATTTTAACTTCACAGTGACACCAAGAGGGGTACTTTTATTATCCCCCTTTAAAGACArs614290TGGTTAAGACCTTGATTAAAAGTAGATTGATTTATTCTGTCAGTAGTTTAGACCTTCCCCCGCTCTTTTAAACATTTGTTTATGGATATTGACACTTACC[G/A]ATTCTTTTTTCAAGGTATGTGGCTATGATATAGCATCCCCTAACGCAGGTCCGCTCTTCAGAGTTCCGATCACTGCAGTTATAGCAGCAAAGTAAGTAACrs686851AGGAAGGAGGCTATACATATAGAAAAAATTTTACACCAGTGTGACAGAATTAGAATCCTCAAAGGGCTTTTACAGACTAGCGTGACGGACCCAATCTAAT[C/T]TGTTGTCTAGGTCTCAGTTTTCTCAGCTGTGAAATGGGGGATCGTGAGATTATCATTCCCATAGGATTGCTGTGGAGATTCAATAAAATAATGGATTAAArs614004TCCATTTCATTTGTACTCAGAGGTAACTAATCTTCATCTCCATTATAACTCCTCTAATAGCACCCAGTTCTTTTCATTCACAGTGTTTCTCATAGTTTTA[G/A]ATTTTTTTATTAGTGCAACACTAGCTGCTGTAACAAATAACTCTCAAATGTTACTGGTTTGACACGATAGCTTTCTCCCTCAGCCATTTAGGAACCCCAGrs558692GCCAATATCTGGTCTCTAAAGACTTTGTCCAATAACTTTCTAAGTCCGTTCCCTTGAATATGTAGATTTGCCTGACTCAGTCAAAAATGTCATCAATCAG[C/A]ATTTTCAGTAAATAATAGTGTAGAGCTTGTTCCCTGAGCCGCAGTGAATGATTGTCCCAACACATGGTGTTCTGAATTATTAGATGAGTAGGAAAGAGTCrs474077TTAAATAAGAAAGAATAATATGTGACATAGGCCATATGTGGCTCACAAAGCATAAAACATTTACCATCTAGCTCTTCATAGATAAAGTTTGCTGACTATG[G/A]ATTAGAAGAATATATATCATATAAAATATAAACAGAGACCTTAGGGAAACGCAAATCAAAACCATAATGAGATATTATTTCATACACACTAGGAAGGCTGrs650616ATTGCCTAAATCAGCGTCAACATGCAGTAAAGGTTGTCTTCAACTGAGCTGTTCTAGTTTTCTCTTCCCCAGCACTGTCATCTAGATTTTCCATTTCAGT[G/A]ATTCCCACCCCTCGGTCTACTAGCAACAACAACTTTCTTGTATCCTTTGAGGAGACGTTAGGGAGAACCATCATTTCACAGTTAAAAGAAAGACAGTCCArs453609CCTTTTATTTGGCCACCTTTTTTCTTGTGGTATTTTCCTCTAATGGAGCTGAGTTTTCTTTGTTCCTGACTTCAAGGGCTTTCATAATGAAGCAGGAAAT[T/C]TTCCCCGACCCCTTCATGGGTAGGAACTGGTGTGCAGGGGCTGGGGCTAGCCGGCCACTTCGGCACCAGCTGAGGCAAACTCTACTCATTGGAACCGGTTrs561470AGTCCAACTGCACGTGCAAGGCTCTGGGGAAATCCCTACCCCGCTGAAATAGGAGCTTGCTGTTAAGTTTCTTGGGTCTCTTGTCACTGATTGGCTGAAA[G/A]TTAAATAAGCTAGTGTTTGAGTGAGGGGCTTTATCGTTTCCTTTAGACCATCTTAAAATAGGGACTCAACCAGCCTGCTTGTCTTAATCATATAAATTTArs356643CCCCACTCATCCAAAAGCTCAGACCAGGTTGCTTGGCCTACCATTCACGACTGCGCCGTCGGTCTCCCCGTGCCTCCCCACCCTATCCTTTCCTTCTCCA[A/G]TTGAGCCCAACTCTGCTGCCAAACTTGACCACCCAGCTTTCCCCTGGGCTGTCCTCTCCACCCCTGACTATGCACCCCTCAAAGCCACCTCATGTTCTTGrs683262TATGTGAATGTGAAGTTGCACACGCCCTGACGGCCCATAGTCTTACTCTTTATATCAGACTTTTCCCAGGCAGAGCATGGAGGTTGTATTTCCTAGTGCA[G/A]TTTAATGACTTGTCAATCTCAGTATTAGAAAACCCAACAGGCATCCTGGTTTCCAGTTTTATTGTAATACCAAACTTTTTTTTTTTTTTTTTGAGACAGArs586030TTTCTTTCTTTTTTTTTTTTTTTGGCCTTCTTATTTGCATTCAGTGAGGAGGTGACACATTGTAGAACATAACCTCCCTTTTTCATTCCATGAATCTAAT[C/T]GTTATTTCTGTGTTTGTGAAAGATAAAGGATTAGCAAGAACGCTTTGCATTAAGTCGACATTTGTAAATGCTTTATAATTATTTATGAAATTATCCTGCArs644818ATCCTGGGCTACAATTTTAGTTCCTTTTGAGAATGAGTCAGCAGATAGTGCTTGAGAGGAGAAAAAAGAAAAAAAAAAGTCTCTCTAAAACTCAAATCAA[A/G]TTCTGTGGGCCTTCAGACAAGTCTGACATTCTTCCATGAGTTGGCTGGGCCGCTGGTCTCAGTTAGGAATACTGACACTTCACCCCAGCAAACCAAAAArs623052GCCTACTTTGTCCAGGTACTATGCTACGGTTTTGGATACAGTATCTTATTTAATCAGCACAGCAACTTCAGGAGGTGCGCCACTAAGCCCAGAGGGATGA[G/A]TTGATGTCTCAAGCTGTCCAGAAACAGCCCTGTAGCCACTGCAGTCCCCTTCTCTACTGTCATATCCTCATTGTATTTTACACTGGTGCTTTCAAACCCTrs1342995TACCTTGTGGAACTGACACCTGTCCTATACCTTAGTCCCAGTGAAATGGGTTCTGATAACATTGCTTGGGTCTTCTCAGCTCCTTCCATGGGACTCATTA[A/T]TTCCCTGTTCCTTTATTGCATATGGATATATCTGCCAGAACCCACCATTGCCCTTGTCATATCTAGATTCCACATTATTAGGCTCAGTCCCTATGCTTGGrs1070036AATTTAAAATCCAAAATGCTCTAAAATATGAAAATTTTTGAGTGCCAGCATGACACTCAAAGAAAATGCTCACTGGAGCATTTCAGATCTTAAGTTTTAC[G/A]ATTAGGGATAATCAACCTAAGTATAATGAAAATATTCTCAAATCCTAAACACTTCTTTTTCCAAGCATCTCAAATAAGGGATACTCCATCTGTATGTTTGrs1003016CTAGGTGAATTTTTCTTCCTCTTAAGCCTTTGGTGACAGGCTAAAGGGTGGGTCCTGGGCCATCCCCTCTTTATACCCCATCTGTCTGTTAATCATTAAT[T/C]GCCAAGAGAGCTCCTCCTGGGATGGGGTGACTCCTTGGCTATGGGGGATGCTGATATCCAGGATGACCTTGAACTTGTTTTTCTTGGCCCCAAACTTTCTrs725849TAAAATGACAACTGCAGTAGAAACAGCTAGGTTCCACAAACAATTGTGTGCTATCTTACACTGTTTAGAATCTTTACTAGAAAGTAGCTTCCTAGCCAAA[T/C]TACATTTTCCAAGCCCATTCCAACTAGTTGGTCCTAGGTGACTAGTTCTGCCAGTGAAATATTAGCAGAAGCAAAGCGCATCGCTTCTGGGATAGGACTCrs1004395CAAGATTTAGGGTTATTTGGTTAAAAAAAAAAAACAGGAAAAAATCCTTGTTCATTTATTACTTTCAGTATCTGGGTAATGAGAGCAGTTTCACCAGTAA[A/T]ATTTACATGAGAGCACAATCAGAGGTAATATTAACGTTTATATGGGCACCAAATTGTGGAATAACCCAAAAGGAGATATGTAATTTCATTAGTACATCTCrs910500CTGAAGTCGAAAATGCATTTGATATACCTAAGCTATCAAGCATCATAGCTTAGCCTCACCTGCCTTCATTGTGCTCAGAGCACTTAAACGTTAGCCTGCA[A/G]TTGGGCAAGTCATATCCAAACAATGCTGACAGCACCGCACACTGTAGGGTATCAGTTGTTTAACTTCGTGACCGTGTGGCTGACTGAGAGCTGTGAATCArs1026791TGGAATAACAATCCTTCTGGCTGCAGAGATTCAGACAAAAACAGGCACCCCCAGATGTCCAAGCACCAAACACAGTGAAAAAGCCAAGATCATTTAAAAT[T/G]GGAGAGTTTGTTTTGGCAGAGAGTGGAAGCAGTACTAAGTAAGAACTGTACACTACTTTGAGCTGTCAACAGAGAATTAATAAAAAGTGAAAACCTGTTGrs748773CAGGAAAACCTTCCCATGTTTCTGATTTTTTTTTCTTTGTCTTTCCTACAACCACCAGTGCAAACACACAACACCCATAATGAATGGAGAACCTGGTTAA[T/C]TGTGTAAACTCCTTCCAAAACAATCAGGTCTTAGCCCTCCCCACTGCATTTGAGAGAGCGACCAAATATGCAACTTAAAGCTAACACACATTGGACGTGGrs1010479TAGCTCACTGCAGCCTCAAACTCTCAGGCTCAAGTGATCCTCCCACTTCAGGCTCCTGAGTAGCTGAGACTACAGTCTCGAGCCACTGCGCTTGACCCCA[G/A]ATTTTTTTTTTATTTTTATTTTTACTTTTTGGGAACAGGGTCTCTCTCTGTCACCTAGGCTGGAGTGCCCTAGTGTGATTGTGGCTCACTGCATCCTGGArs1259733AAGGCAGCTCTGGGGAAATTCGTCTGTGTAACTGGGGTGCTATGCAGGCCTGTCTGTTTGACTGTCATGCAGGTCTGTCTGTGTGATCATCAGGGAGAAT[C/T]GGCCGGCCACATTTTCAATCTTTCTCCTCTTCTGGAAAAAATAACCACTCAGTCTTTAGCGTCAGCTCACCCTTCAGGGTTTGACACCTGGAGCAGCAGTrs726395GAACAAGATCTCAAGGAAACACAGGAAATTCCTGAAACAGCAAAACCCCCTATGTGAAGACGTCTGTTATTCCTTTGTGTATCAGGAGCATCACTGTTAA[T/G]TAACTGTTGATACACTTAAAACAGCTTGGTTGGCAATGTCCTTTTTGAATAAAGAAGAGCCAGGTGTGTTTTTCAGGATGAACTAGACAATGCTGTCACArs1335075AGAAGCTGAGCCCTGACTTCAGGACACATGAGGTGCAGGACCAGAGTCGCCCCTAATCAGCTAGGCAGATTGTGCTGTGTTATTCCACTTATGAAACAGC[A/C]ATTGCAGATCCTATTGTGGCCTTCTCGGGAGAATAACTGCTTCAGGCTTTGGGGATAGTTAATTTTATTGTAAAATTTACACGGATTGGGCATAAGGTTGrs1054067GGAGTGGAAGCTCTCTAAGGGATACGAAATCTGATTTTATCCAAAGAAATCACAGCAGCCTTTGAAAGACACATTCAGATTTTCAAACAAACAACAGACA[A/T]TTCAAAACCAGGCGCTCTGCTTATTCACTGAAACCGTAAGCTGCTGAAACTCAGGGAAAAGCTTAAGAAACTGGTCTTAAAGCTTCAGGCCAAACAATGCrs1029176TGGGTTTTAGCTGAATCTGTGATAAAGTTAATGGATATACCTTTGAGAGACACTGTCTCACTCATAATAGTTGCTAACAATAGAAACCATTGTAGCTAAT[A/T]CATGTAGTTGAAAGCATATTTATGACTGCTAGAGATTCAGGCTTAGAGGCTATGCAGCCAAAAGTCTCCCAACTATATCATAGAAATGCACCAGCAAAGArs880385CTAAAGTGCCATCCTCACGTGAAGGGTTGTTGTTGCTGACAGCGTTTGTGAAAGGCCACAAAGCTGTTGACATCTGCTGCTTGGCACTGGCTGGGAAAAA[T/G]TTCAGAGGGTCCCATACTCATGCATGTGAACTTCTGGGATTTCTCATTCTGGGAACATTATCAAGAGGCACTAGTTGACTCCCTTTGTCCCCAAGGGAGCrs1458207CATTGCTCTACAGCCTGGGGGACAAGAGCAAAACTTCACCTCAATAAATAAATAAATAAATAAATATTATATATTGGCTATTCTTAAATCTATATATCCA[A/G]TTGGACTCCCACTTAGATATTTGAGAAATATCATATACATACATGTCTAAACAGAACTAATGTTATTCCACTATACCCCTAAACCCACTTTTACCCCAGTrs1376827ATCAAAAATAAATAAATAAAACAAGAGGAAATTGATTTTGTGGAGAGCATGGACACATTTGTTGTCCATAGGGAACTAACAATAACTTCCAGTGACATCA[G/A]TTCAAGAAAAAAAAATAGCAGCAGGGAGTGAGAATGTCATCTGTCAACCCCGAAAATGATTTTGGTTAAAATAATGACAACAACAACAAAACTAAATATCrs2063506AAGTATGGGGAAAGAAAAGTTAAAAAGAGGAGGCAGAAATAGAAACTATTTTAGGTGAGATAAACTCTTTAAGGACTCCCTACTCATTCAAGTCTTCTAA[A/G]TTCCTTCCCCTGAGGCTTTATTTCTGAAATGGGATCCTTGATTCTCATAAACCCTTAAACTGGAGGATACTTTGGTTTCCAGTATTATGGGCCCAATTCTrs1593443CCTTTCTATGAAAATTTCAGAGTGAGCTTGGCAATCTCTATAGAATCACTTACTGGAGATTTGATAGGAAGTGCATTAAACATATATAAATGTGAAGAGA[G/A]TTGATACCTTTACTATATTGAGTTTTCGAGTTCAATGAGCATGGTATGTTTCTCCAAGTATTTAAATCATCTTTGACTGATTTCATTAGCATTTTTAATTrs2401505AGGTAGATGACCAGCGATGCTGTTTACTACATCCTACGCATGCTGAGATAGATGAGTGGGACACCATCTCGGTAGGAAAGAGTTTGACAGGAAGAAGAAA[T/C]TGCAGAGTCCCTGCCTGTCCCAGAGAACCTAACAAACTGATCTATATGGAAAGGATTTTCAGCAAACATGCACAAACACAGTTAATATGCTGGAAGAAGArs2367059AAGGAAGGAAGAAATGCTAAAGCTACAGTTTTATGACTTTTGTTTGCATGATTTCTGCTCAAATAGGCTGGAGGAATGGGATGATGACTATATTCTAACA[A/T]TTAAAATGCTGTCAATACTAGAACTGTAATACCACACTCCTTTAAACAAGAAAAAGCCACATTTCTGTTTTTGTGGTTGACTACATGATTTAAGTTTTTGrs1367452CCACAGCAAGAGTTAGAGACACAGCCAGGACCAGAACTTACTGAAAAGCAATAATCCAAGGTCTGACTTGGGTTCTGAGGATTTGCTCAGTTATAGATGT[C/A]ATTCTCGATTTGGCCTGAAGAGGGCAGACACTTGCCATTCATGGAGGTGAAGAGCTTGTTTTTAAGGCTGGACTAACATCTCCCAATAAGGTCATTTGGTrs2241491AGGACCAAGGATACAGTCCAGATGTGTCATATAAAAATAAGACCCTTGCACCCGACTGAGTCCACAGCATATTAAGTTTATATATCCATCCATAGTTATA[T/A]TTGGGAGAGCTTTGGGAGAAATAATGGTATTCATTATAAGAGGCTTTTAAAAAAGTATTTCAGCTAAAAAAAATTGAGGCATGCTCTTTAAATTGTTTACrs2007475CCTGCCTGTTTCCTGGACAGCCGCAGCCTGTACATAGCTTGGGCTGAATGTTAGGTAGAAAGATTTGGGACAAAGGAGAATCAGCGTTTCACGTTCAAAA[T/C]TGAACTGGGAAGCTGTTTTGCTTTTAAGTTAAATCAATGAATGAATAGATCGAAACAGAGAGAGAGAGGAAAGCATTAAAAAGTCCCCTGAAATTCATTGrs1372688TTTCCAGTAATGAACCATATAGACATCATTTACTCCCTTATATAATACAGGGATAATACACTTATACCTAATGCTTGTTAAATGTGTGTTCCATCATCTA[T/A]TTATGAGAAAATGTCAGACAAAACTAAACTGAGGGATATTCTACAAAACAAATTGGTAAGTACTTTTCAAATATGTCAAGCTCTGTCTCAATGTCTCAGTrs2304748GGAGGGGGCTGCATACTTCTCCAGGGCTTTTGCTACCTGCCCCAGTGTCTCCGGCATTCCGCGGAGCCGTGCACCAGTCCCCTCAAATAACCTATCAAAT[T/C]CTGTGGCTTCTATTAGATCACCGAGGTCTTTAAGAACTGCATCTTTTACATTGCCTCGTCGTGCCTCCGGGCGGGCAAGACAGTGTAAAATCTTGGAGAArs2207800GCACGAGCTAAGGGGGCGGATGGATTCTGAAGAAGTATTTCCACTACCAAGAAAGGCAATGTTGGGAAGAACCTTTTGGTCAACTTGGTCAACTTCTCCA[G/A]TTCCAGTAGAGCAATCCAGCTTCTTTTGTCATCTCCTCTTTATCTTTCATCCAGAGAGCCCACATAAGCACTGTTTAAATTTCTCTTGACTTGTCTTACTrs2373814GAATGTCACAAAAACCCAGAATGTCACAGTATTGTTTTCTTCTTGCTGGTGTCCTATCCTCTCTCCTAACACCAGCCACCAAAGCTGATTTTTAAAAAAT[G/T]CCATGATTTCTCTTGTTTACAAGAAGCTGTTTCCTATACCCTATTCTTGAAGGATAAAGAAATAGTCATTCAAAAGAAATATCTGGCTTTTCACAGTGTTrs1543513ATATAACATTTGGCACTTATAGAAAAATATAACAAGGAAAAATTATTCCACATTCAGAGACAACTAGTTTTTTCATATGTATGTGTAAAAGTACATATAA[C/A]TTTATTTTGCCTGCTTTAAACAGAACTTACTCTACCAGTCTTCTGAAAGCAAAATTTGTGTTCTACATAGGCAGTTAGTTCAGTAATCCAAGTTTTTATTrs1904161TCATTTGTTGAAACAAACTTTATTTGTTTCTCTGTTTAGAACTATATCTGTGTATTTATTTTAATAATATTATAAGCATCCATGGACCTACCACCCAAAT[T/C]GAGAGCTAGCTCTCTTTTTTGGTAACTACATCTGAGCACATTTACCACCTGCCTTCCTCCACCTGGGTAACCATCAGTCCACTTTCTACATTCCTCAACCrs1444647ATATTATGTACAACTATATGAATTTAAGATGATTTCTTTATCTAGTCTCCCATCTATGATTTCCAGTTTTTGTCTACTTCAAATCATGCCTCTATTGACA[T/A]TTTTGTGTGTGTCTCCAGATATGCATGTAGAGATAGTTCTCTGTGATATGTTCCTGGAAATAGAATTGGTGAGTTGTAGTGTACTCATAACCTTAACTTTrs1420562CCAGAATTCAAATCAATGCTAACCCCAGAGCTCGCATTTTTAAAACCCTACACCACACTGGCTCCATTCAAGTGTTGATATGAGCCTCTGAGACTGAAAT[T/C]AAAACAAGCAGATTTCGTTCAACTTATTTAACACAAAACCTTCTTTTGTAGGATCTCACGAACTTCAGCTTGCACACATCCCGTGTTGGAAGATGCCAGCrs1850422AATATATAATACGTGAGAGGTGATAAGCGTTATGTAGAAATTAAGCAGGATTAAGAGGTATAGTGGAAATGACTGTAGGTTAAGAGGGAAAGGGTGAAAA[T/A]TCAGAGATGTTCTTTCAGTCAAGAACAAGGCAAGTATTAAATAAACTTATACTTAGGTCTACTGTTCTTAATATGTTCCAGCTCCTGCATGGTCCCTGTTrs1916803TGAGCAGGTTCCTAGCTCGGTGCCTTGCACAAAACTGGAGCTTAATGTTTGTTGATGAGGTGAAGAGGGGGATACTTATCAGGGGCCACATTCATGGGAA[T/G]TGGATACTGAGACAGAAGTGGGTGAGTCAAAGGTTTATTGGGCAGTGACACTTGTGACAGTCCAGGGCATGAAGCAGAAGCAGGATCAGGCGGGAGGAGCrs2451984TCTTGTTTTTCTGGAGACCTTGTTATTCAGCCTTTTCTTTAATCCAGGGAGCTCTTCCATATTTTTCAAATATCCTGAGTTTTTTGTTTGTTTTTTACTT[C/A]ATTTAGCCGGAGTGTGTCTCTATTGTTTGCCAATGATCTAAAGGATATGTTCGTTTAGTATTTTGACAAATACCTCTAATTGTCTTCCAATCGGAGGTAGrs2462049TTAAAAAGTTCTCTATAGAGTGGGATTTTTTAATAATACGAAGTTGGGGAAGGGGAAGGTGTTTGTCTCATATTTACTTTCTGCAGCTCATATTCTGCAA[G/T]TAATATTCTTGCTCCTTTCAAACTGTACCAAAACACCCTCATTAAGCAGTCAAGCTATAACCACAACAGCATCACCACACCCTCAAGAACAGTTGAGTTTrs1503660GAATTAAGTAGAACCACTGTCACCACAACTTAACAACTATGATGTGCCAAGAGGTTTCATAGACTTTATAGTCTGATTAGGCCTAAGAGCTGGCTTTTAG[A/T]ATTTACTATCTGTTATTGAAACTGCTTCCTTGACTGGTATATCTAACAGTTTGGTCAGATAACTTCATCCTAAAATTACAGAAGTGAGAAGGGGTTAAAGrs2092797AAAAATTGAAGGTCTGCAGCAACTCCATTTTGAGCAAGTATATTAGCACTATATTTCCAACAGTGTATGCTTGCTTCATAACTCTGTCACGTTTCAGTAA[C/T]TCTTGCAATATTTCAAACCTTTTCATTATTATTATATCTGTTACGATACTGTTTTGTTTGTTTGTTTTATTTTGTTTTGAGACGGAGTCTCGCTCTGTTArs1401454AAAGGGTACAAAGTTGCAGGTATGTAGGATGAATAAGTCTATAGCTCTACTCTACAACATAAAAACTGAAGTTGATAATATTGTGCTGCATGCTGTAAAT[C/T]TGCTAGGAGAGTATATTTTAGGTGCTTTTACCACACACACAGAACAAGGTAACTATGTGAGGTGATGCATATGTTAATTTGCTTGACTAGTAATCATTTCrs2427102CACTCTTCCCCCACTCCCTATTGCAAGAATCCCAGCCTGACCCTGGCCACCTCTGGCCAGGGATTGTATTCGAAGACTGTCAGGAAGCTCTGGAATCAAT[G/T]GAGCTGGGGGACCCCAGCTGAACAATATCCAGGAACCAAGAGGCCTGTGGAGAGCCAGGCAGGGCCCCGGCCATCCCCAGGCAGGACAGCATCAGTGCTCrs1912619AAGCTGCAGCACACTCATTCCACTTTGAATATAATGGAAGAAGAAATGCCCATCCTTCACTAACTTGAACTACAAGATTATTTTCCACCCTCAGGAGGAA[C/T]TGGTCTTTTCCCACCACTGATGGGCCACCACTGTTGCAGGATTTAAGTGTTACCTCGGAAATACCAAAAAGATAGTTCTATTACAATGTTGTATCCTATArs1378933TCACATACTTATTGTGTGCCAGTTGCTGTAGTGGGCACTGTAATACAAAGGTGAATGAGGCACAGGCACAGGCTTTAACTTTCAATGGAGAAGACAGAAT[T/C]GTAAGCAATAGTTGTGATACCATGTGTGGAGAGGATGGTAGAGACAAGAACAGGATGTCATGGGATACCTGGCCCAGAGGGCCACTCAACCCAGCTGAGGrs2034877GTCAGGTATTTTGCAAAATGTCCTTCAATTTGGGGTCATCTGATATTTTCCTATGATTAGATTTAGGTTATGCATTTTGGGAAATAATACCACATCCAAT[T/C]TGTAATCTGGATTTTGTGAATAGGTAGACATTTAGTTCACTTTCATCCTGACTTCCCACAGGTAACATGCCTCCTTGTATTATCTCCCAGTCCTTTGCCCrs1548605AGTGCACGGCCAATGAGGCCTTGTGAAGTCAAGTTCCAGTGTGGAATTTGGATGGTGATAATGAGAGATTGAGCTTCAGTCCCCTAGTGTAATAGGAAAT[G/T]CCACAACGAGATATAAAATCCTTACATGAAGTTTCCCTATCTACACAAGACTGAATCGAGGCTATTTCAGTTCGTGTTGCTGAATGTTCTCTCTTGGTTTrs1540885TCAATGATAACTTCCTCACTGCTCTGCAAAACAATATGCTCTCTTTGGGAGAAAATGAAGAAGAATCAGAGCCAGCTAGCTAGCTAGCTAGCTAACGGGC[G/A]ATTGTTCAAAACCTGGGGGGCACATGGGAGTATGTCATAAAGTCATGTCACCTGCCAGCTTGCCAGCTTTCTAAGTAGGGTGAAAGGATTAAGTAAGAACrs1870836AGGATGGACTGTAAATCCAATGAGAAGTGTTCTTATAAGAGAAAGGAAGGGAGGGAAGAAAACATAAGAGGAGAAGGTGATGTGAAGATGGAGGTAGAAA[C/T]TGGAGTGAAGCATCTGCAAGAAAAGGAACTCCAGGGATTGCCAGCAGCCACCAGAAGCTACAAGACGCATGGAATGAATTCTCCCTCGGAGCCTCTAGAArs1822243CTCTACTTTCTGTCAACCTGGAGAAAGTCACTTAACCTTTCAGCACCTGTCTCACAGTGAAAGTGAACAGTTTAGGTCAAGAATGCTTTCAGCTCCAAAA[C/T]TCTAAGTCCAATACGATCACAGAAAAATAAAGTGGCTACATATACGGGTGCACACACACACAGAGGTTTGTCTGTGCCAAGAGAGCTCCACAGGAGTCTGrs1536069GGAGTAGATGCCTGAGGTTGTCAGCAAATATTGAAGATTTTTATTTCCAGGGGGTTAGAAACTTCACGTAGCTTCTCTGCTCTGCACACATAAGGAGTAA[G/T]TGGATTATTTTCCCTGAGTCAGTAAGGTTTTCGGTGTCACATAATCTCAGGGGAACAAATGCAATTGCTTAGATCTCAATATACTCCTCAGATGGCAACTrs1363267AGATGATGTAGCTGGACATTTAAAAAAGCACAGTAGTACATGCAGGGTCACATCACAGATTGAAATGAAAAAAGTCCTTGTTGTCATTTTTATTTCACCA[A/G]TTGGAATAAGTTTTCAACTTGTGAAAAGTGCTGCACAAATCCTGGAAACTGAAATTCTTTACTAAAGCACAGGGAAGTGCAGGGCAATCAATGGCAATATrs1797700CTGGTCTTAGAACTCCTGACCTCAGGTGATTTGCCCCGCTCAGCCTCCCAAAGTGCTGGGATTACAGGAGTGAGCCACCACGCCTGGCCAGAACTAATCAA[C/T]TATGTTTTTGTTGCATCTTTGCCTGTCTCTCCCACTGGGCTATAAGCTCCTTGAGACCGGGAATTGTGGCTTTGTCTTATATACTTCTGCCTAACACAATrs2435556AAGAAGATGCCATCCAGGACTTTCATAGCAAGAGAGGGGAAACCAATGCCTGGCTTCAAAGCTTCAAAGGACAGGCTGACTCATTTGTTAGGGGCTCATG[C/A]ATTTGGCGATTTTAAGTTGAAACCAATGCTCATTTACCATTCTGAAAATCCTAGGGTCCTTTAGAACTGGTCAAGTCTACCCTGCCTGTGCTTTAGAAATrs2126316TTCTATGAGGCCAGTATCATCCTGATACCAAAACCTGCCAGAAATACAACAAAAAAATAGAACACTTTAGGCCAATATCCTTAATGAATATCGATGTGAA[A/G]ATTGTCAACAAAATACTAGCAAACTGAATCCAGCAGCACATCAAAAAGCTTATCCACCACAATCAAGTAGGCTTCATCCCGGGGTTGCAAGGTTAGTTCArs1418136GTACAATCTCCAAGACATTTTAATTTTACCCTGTCTTTTATCTGACAGAGTTACCTGCATATTTTCTTATATATCGTCACCTTATATTTTCAGAAAAATA[A/T]TTGTACTTCAATAGAAATCTCTATGCATGCTCTGTAGCATGCTCCAGGTTACCTGAATCTGATTTTATGGAAACTATTTTATAAGTCCGTAAGTCATAGArs1432865CCATGCTACTTCTCCAGTTCACAAGCTGCAGCCACACAAAACCCAGACCTGCCTCGGGGCCTTTGCACTTACTGCTTCCCTTAAGATTCTCACATGAATA[A/G]TTCCTTCTTGTCATTCAGTTTTCAGCTTAAATGTCACCTCCTGAGCTCCTGTTTGGAGTAGACTTCCTGTCAATTCCCCTCTTTATCACTTTGCCCTATTrs1885121TTTCCATTTTTCATTAGCCCCACTGTCCACATGCTCTTGACCATTCTCAGAGTCGGGATCTGACCATGACTCTAGTGACCTTCAATATATATAATCATAA[G/A]TTGGTGTCCTTTGTCTTATAGTTGTTTCCTGAAGAATCGTCTCAAATGTATACAAATCCTGGCATTTAATTGTGGGAATGGATCTGCTACTGTGCACAAArs1720839TTAAAGGTATTGAAAATCCACATTGGCCAAGAGCTTATTCTATTTTCAAGTAGAGATGTTGCAAAGATGCAAGATTCTCAAAATATAGTGAAAGGTTGAA[A/G]ATTAAAAGACTTATGCTTTCATCATCTTTTCTTTATCATAACATGCATAAATGTTCTTATAGACTGATATGACAGGTCCTTCAGTACCATATGCTCACAGrs2030926GAAAGTTCTTCATTTTACGGGCTGTGAAAAGGGGGCATCACAAGTGACCAGTCCAAGGGCACACAAATGGATAGGGATAGACACAGGACAAGAAACCAAA[T/C]TTCCTCAATGCCAACCAGTGCTTCTCATACCCTGCTCACCTTTAACTACAAGATGTCAAACATCAAGATAAAAATAGCATGCTTGGCCGGGTGCGGTGGCrs2247858ATTGGTCTTGAAACAGTTGATGCTTTGCCCACATGAAAAAAAAAGCAGTGGTATCAATAGCCAACACCACTTAGCTAAATGACCTTGTTCCTAGAAACAA[C/T]TGCTTAACACTATTGTGTACAGAGTCCTGGACATACTGTAACTTTTCTGATTATCACAATGCACCAAAATACATCATCTACTAATGCACTGTATATAAATrs1597205TAATGTTAGCTGTGGTTTTGTCATATATGGCCTTTATTATGTTGAGGCACTTTTTTCTGTTCCTAGTTTGTTGAGACTTTTCTTTTTGTAATCAATAAAT[G/T]ATATCAGATTTTGTCAAATGCTTATTCTGCATCTATTGAGATTATTGTGTAACTTTTATTCCTTATTCTGTGATTATGGTGTATCACATTAACTAATTTTrs1346718ATAATGTATACATAGGTTTTCTGAGGGTGTAAAAGTTCATGTGATTCACGTCTTGGAAAGTGGAGGCTGAAGAATGCTTTCCCATTGGGTTAGCAGCTGA[A/G]TTGGTCTGAAGAGGATAGTCAAGGGAAAGGCTTGCATCCATACAAAAGAAAAAGTAATAAACCGAGATCACAAAGTATATGAGGGGCTTCCTGACACCTArs1514424TAATTCTTCTACTTGCCTGATACTCATGGCATATCAACATTACTTTGATGAAAAACATTAAATCTCTTTGGATTAAATGCCTGCAGGTAATATCAAGTAT[G/A]ATTTACCTCTCACAAGCCTATTACACATGTTTAGGAAAGACGTTAAAAAAACAGTATTTCGAACAATTAATGCTGTAGTTGTGTTAACCTGTGTAACTGArs2168524AGAACTCAAAACAGAAGCAAGCAAGCCCTCAAAGGAACTGAGAAAATTTCTCCCCACTTTGTTCTGAGGGGTCTCAGCTACTCTGGTATTTAAAATAAAT[G/T]GGTTTTGAAAAATAGGTTACTGCCCTTTAGTTGATGACTAAAACAGAAGCCAAGAAGTGTGCAAATTGCAAACTGACATGCATGAGCCAAACATATTCTCrs1910369GACTCCATCCCCTACCACCATGTCACAATGTGATAGAAACCACTGGGTAAACATATTTCAGATAATAGTCCAAGGGGCTTGAATAGCTAGATACCCAAAT[C/T]CCCTTTTATCTTTATCTTGAACTGCGTCTGGCCTCCAGATCTCTAGCTAGGTAATCAAAGTGGCTGGTTTTTATTCTTTTCATGTTGCAACACCTAGAGArs1445496ACACTAACTTACCATAATAACATCTTTTAAACTATTTTCCATATCATTAAGTATCAAGTGTTGTTACCCTGTAGTAGTACAGAAGTAACAGTAAACTAGC[A/G]ATTAATAGAAAAAGCTAGATTCCTGAAGGTTATGGCATTTAGAAAGATCTTAATTGTTCACAATGGTAAAACTAGGAAACAAAAGAATTCTATAGCCTCArs1462685TCATGATAACACCAAAAGGCTGTGGCAGAGTTATCTTATACATTCACACAATTCTTATAATAGGGGCTGCCAGCATTCTGAATGGAATGTAATAGATATT[A/G]ATTTGCACACCCAGGTGTGAATATTGTGATTTCCTTTTACCACCTTCCAGCTTGGAAATAAATGAGCTTCTACTGTTTGTTGTTGCTCCTTTCCTCATTCrs2298810AATTTCTCATTCTGGGTACCTCATATTGCAAAAGAGCCTGTGGCCTCTGAGCTGACTTGGTCCAGTAGGAAAACAGGGAAAACAGTTCGTATTTCAGAAT[T/C]GACTGTCACAGCCTTGAGACCTGAAATGTAGCCCCCATCCATGAGTCAGTGAAATATCTGTATTTCTTAATTTTCTTTCCTTAAAACCACACTCTCCCTGrs2191076TCTTTGCCATTGTGAAGAGTGTGGCAATAAACATTTGCGTTCATATGTCTTTATAGTAGAATGATTTATATTCCTCTGGATATATGTCCAGTAATGAAAT[T/A]CCTGGGTCAAACAGTATTTCTGTTTTTAGCTTTTGTGGAGGCACCATACCGCTTTCCACAATGGATAAACTAATTTACACTCCCACCAACAGTGTATAAArs1439047TTTCAAATACTGCAAAAATGTTTCAGCAAGCATCCTTATTCATGCTTTTTACACATATGTACAAGTGCAAAGTTTTTTCTAGAATACACAGAAAGAAGCAG[A/G]ATTGTTGGTTTATGTGGTTTGCACATGAAAAAATGGCTGTATCTATTTATGCCCACCCTAACAAGGTATACCTTGATATTAGCAAACTTGTTAGTGTTTGrs1904185AATTTTAAAATTTAATTTCAAAATAGGAAATACATAGGAACAAATCTGACAAAGATATAAAAGATGTGTACCCTAACATGTGTAAAACATTGCTGCACAA[C/A]TTAAAGACCTAAATGAATGGGAAGATATACTATACTGTGTTCATGGGTCAGAAGCTTCAATATGATTAAGCTGTCAGTTCTCCCATATCAAAATCCTAGTrs2322301TCAGTTGATGTCAGCTCCATCTTCTCAGGTGTTCAGAGGAAAAGACTCAGAGTCATTCTCATTCTCTTTTTCCTCCTGTACCCTGCAATCTGTCTGGAAA[T/C]TATATTAGTCCTATCATAAAAAATGATTCCAGACTCTAACCACATGTATCTATCTCCACTGCTACCCCAAGCAGATTCAGGTCCTCCTCTCCTCCACCTTrs1789529GATTTTGCCAGAGATCATTCTTTGATGTGGGAAGTGGTCCTGTGCATTAGAGGATGCTTCGCAGCAACCTGGCCTCTGTTAACTATTTGTAAGTAGCAAA[C/T]TCCTCACTCCTTGTGACAATAAAAAATGTCTCCACACATCACCAAATGTCTTATATGGGCAAAATTGGCCCCAGTGGCAACTACTTCTTCAAAACTGCCCrs4489023CAACATGTGTTGAGAATACTTTTGCAAATGTTAAAGTCAACATGGCTATAACAAGCCCAAGTTCTCCAGGAGGAATGCATGCATTTAAAATGGAATCAAA[A/G]TTTATAGAGTACAATAATAAGAGCCCTCTTACTTACATTTTCATTTAATCACATGTATATGGCCATCTTGTCCATTTTGAGGTTGGGCTTTAGGGAAAGCrs7266163CAAATTTCTTACCCACAAAGTTCATGAGAAATAATATAATATTTGTTGTCTTTTCGCTAAGATTTGTGTTGATCTGTCACATGGCAATATAAATGACCAA[C/T]TGAGCTATTTTCTCAAACTTCAGGGCTATTTGTTCTCATTGAAAGATTATATACAATACTCAGGAAACTTCATATAATCATCTATGTGATGTTTCTAATTrs2865878AGTACATTCCCTTTTAGGGTCCCTGTGCTGTGAATCTCATAATTGCTCCAGATTGTGGCTGTGCTGTCCTCCTGGGTCTCAGAGGGGGGACGATGCAGAA[T/C]TGAGTCCCTCCCCAGGATCCAAGACAGATACGAATGTTAGAAAGAAATAACCCTTTGTTATTTTGAACCAGGACGATGTGGTGCTGCTTATGACCCACTGrs7320201TCCAGAACATCCTGGGTTCAGGCATATAGGTTGTAGAACAGTAAGATTTCTGTTCAGATTTCTTTTGTTTATGCTCATTTATAGAAGCAGTCTTTTTTTT[A/T]ATTTCAGTAGGTTTTTGGGGAACAGGTGGTGCTTGGTTACATGAAAAAGTTCTTTACTGGTGATTTCTGAGATTTTCATGCCCCTATCACCCGACCAGTGrs4764597CGACTTGCTGTGCCTTCTGGCATCCGCTTCCCAATCAGAAACCTCACACATGTCTGCAAAGCTCCCCCCAGCCAGATCCTCCAGCTCATCTTCCTCTGAA[C/T]TGTGTTAGTTGTACATATGGAAATCCAGAGAGCCTCCAAGGATTAGAGTCCACGTCTTTTTTTATTTGGAACTCTTACCTGCCGACCCATCATCAAGGACrs4399565ATTTTGTGAGGATGTAAGCAAACTAAGAAAATGTCAGATATGTCCAGCTACTAGTCTAAAGTGTTGACCCCAGTGTGGGGGGCAGAGAGGGAGCATGTAA[G/A]TTGTCCTCATCTCTAGAGCAGCTTCACAGAAATCCAGAGGTTCTTTTAGCTCTGACACTCTCTAACTCTGGGAATCACTAAGTCAATGGAGTTCAGAGGGrs6542638TTATACGAGGCTGAGTCCCCCAGACCTGGGCTACTTGGGTCTAGGAAATAGAGGCTGAAAGTACTAATGGCTCAGTTTAAAGTCACTGCCAGTGACCTAA[T/C]TGGGAGAGTTTTTTATTTTCTTTCCTGAAACTCTAGTCTTTGCAGGGTTATAGATTCTAGCTCCCAAAGGGGAAAATATTTCACAGGGGACACTATAGGArs4953843TTATCAGAAGAAAGTGTAAGAATTAAAGTTCTCATTACAAAAGCTTTGCGCATCAGGCATTTTATACTAGGAATGCTCAAAATCTAAAGCAGAGATAAAT[C/T]ACATTAATATGGTTGAAAGCAAGGGTCCTGTATGTATTCTTGAAGAGAGGGACTCTATCCTGCAGTAGATTATAAAAATTTAAGAGCATCCTTCTCCTTCrs7810506TACATCTGCTTTAGCACCCAAGCTCTTGCTTGGTGAAAAATTAATAGTAAACATTCATCTTTTGAGCATCTTCAAATATCCCCTTTAGAATGACATTCAA[T/C]TATTAGGTCAGTAACCCCAAGAGAAAACGGTTGTTTGAGTGTATATACTGTATTACAAAATAAGGGGTGAATTCAAAGGAAAACATAAGATGCAATTCGTrs4708590TGGTAAGAGAATCCGCACCTGAAGAGACTGGGAATGAATGGAAATTTTCCTCCCAAGAGAAGGGCTTTGCATCCTCCAGGGCCAACTGGATAGCCGTGGA[A/C]ATTGGCTGTGCAGTGGGCTTCTTCTCGCAGCTCTGCAGTCTTCTGGGGCTGTCAGCCACGATCACCTGCGTATGCCTGATGATTGCCACTCACAGGGAGArs3128688AAGCCTGAAGTTTAAACTACCATTTTGAGATCTACCCTAGAGATTTACTCAACTCTCTGGGTTATTTCTCATGTGTACAGAACATATACTTGTACATGCA[A/T]TTCAACTTCTGTCCATTTTCCTCTTGATAATCTGTTTTATTCTTCCCGGGAGTCTCAGCTAAGAACTCATGAAGTGGAGAATATTATTTTTCCTCACCTArs7689368TAATCTATTCATAAGAAAAATATCTATGAACCCAATTGAGAGACATTCTACAAAATACCTGACTAATACTCAGGTTGAGGTTATAAAAATAATGTAAAAA[A/C]TTTTCACAATCTAGAGGATCCTGTGGAAACATGGCAACTAAATATAATGTAGTATCCTGGATAGGATAACGGGACAGAAAAATAACATTAGTAAAAACTArs6962207TGACTTCCCCTGTGGCATGCCTGCAAGGAAAATACATTGACCAAAGGATTTGAGTGATAGGTCCTCTCTGCAGTCATTTTTTAAAATGGAAATCAATAAA[C/T]TCGTATTCTTATTTTGTGTGTTAGTTTTGTGAGCTTGGTTAATGTGATTTCCCCTATTTGTACTGACTGACATACCATCATCATCAACCTGCAAAGGTTGrs2647415AGTTTGTATTTGACTCAAATACAATGTGAGTGGCTTTGTGGAATTAAGATATAGAGATAGATTTGCTACGATTCAGTAATGAGTACAAGGTATAAGAGCA[A/G]ATTACCATCATAGTGTCTTTTCTTGCTCACGTCCATTTACTCAACAAGACTTATTGAACATAAGGCACTGGTCCAGATTTTTCCAAGGACCAGTTAAGATrs6766358CTGTCAGACATTCAAAGAAGAATCAGTACCAATTCTATTGACACTATTCCACATGCTAGAGAAAGAGGGAATCCTCCCTAAATCATTCTATGAAGCCAAT[T/A]TCAGCCTAATATGAAAATCAGGGAAGGACATAACAAAAAAAGAAAACTACAGACAATATCCCTGATGAACATAGATGCAGAAATCCTCAATAAAATACTArs4894467AGCACGCTTGTACTGTATTTCTCTTGGCCCCTTTCATCTAGAATTTATGCAAGAGAAGGTCCTGTTAGTAGGGGTTAAACATTTGGATTCAGCTATTCCT[A/G]ATTGCATTTTAGTTATTACATCATGTAACCCAATACATTTCTTTTGTTGTTGTTACTCTTTTTTGCTTGATTCATTTTTAATGTTTCCTTTTGTATTAATrs7818415CTGGCATATTTTAAACACTCAAAAATATTTTCTGTAAAAATAGTCCTTGTTAGACCTCCACCTATGAAACCATATCACAGTTGTTGGGTTTTTTTGTCCA[T/A]TTGTTTATTTTAGTTAAGCTCTCATTTGTTTTTAAGAAAAACTCTAGGTCTTATAACTCCTCATTAAATCTATCCTACAGCTCCTCTTGATGTCCAGTTArs2928668AACAGCATGACTCTGGCCGACCGCCTGGCTTCTTTCTAGTCTTCCTTCTGTACTTTGTGACCTTGAGGCAAGTCATTTGGTCTCTGTGCCTCAGTTTCCC[A/C]ATTTGTGGAATGGGGATAACTGGTTGCTAATATTGCTGTTTTTATTGTTATAATTATTTTGTAAATAGAAGGGTTGAAGGTTCAGGGAAGCAAGTTGATTrs2993531ACACAATTAATGCCAACATTTGTACTTACATTTTCCATTTTATGAATTTAAGTCTTGTTTCCCCAACATAAGGTAAACCTTCTTGGAAAACACACCTTGT[A/C]ATTTACACTTTCCTGTATCTCTCAGTGTTTATATAAGTTGATCAGTTTTTCCTTCAAAAATTTTTCTTGGGAAGCCAAATTACTAAAAGGGATGTACTTTrs6941784ATTGCATTTTATATATATGTAATGTTCCACAAAATGTTATATAAATGACATTTACCCACAAAGGTAAGAATAAGAGGAATGAAGAGATTAAAATAGATAA[C/T]TCTAAGTTTCTCTCCAATGTCAGGGACTAGGCCTTTTACATCTTCATGCCCGGTCACTGGCACATACTGAACTTTCATATACTTTTCTGCAGCATGATTGrs4680921ATATGGATGTGGGTGAACATGACAGTTTCAACTATAATTGCCAAGCAACACTATGGTATTATCTGTATTGGTTGACACCTTTTAGTCTAAGGAGAGAAAT[C/T]GCCAAGTGGCACAGTTCACTTGGTCTTAAAGAGACATGAGTTGGTCTTCACCTTGACACAGAGCCTTGCAAGATTAGCCAGTCAGCTCTGTGAAAGCAGTrs4716945CTTTGTAAGTGAGCTTGTGAGGTTGCAGGATCTTAGGATCTTGCCTCAGAACTTCGAAGCAGCATGAAGCATCTAAGCACAGCTCTGTGGAGCACAGAAA[A/G]TTTGAAAAGAGCACCTCATTCTTGGCTCCTGAGGAAATGGCATTTGTTTGCGTCTGTAAGGAAACCACACAGGGCAGTGTTTACAAGTATTTCGATTAAArs4897019TCAAAACTTCCTCCCAACTTCTAAAAGTTCAGCCAAGAAAAAAGAATTTAGAAGGCAAGGCAGGGAAGATAAAAACCAGACATTCTGTTTCCCAAAAAAT[T/C]GACTTCCTTCTTCCCTTTGAAAATCTCCTTTTCTGCAAAATATTGCCCTATTGTGGGAGATTTTGCTCTTCTACCTTAAAAACTCCTGGAGTCCTCCTTGrs4667489AATTTTCAGAATTAATTTAATTCTGTGCCTGAATTATTTTAATTTTCATGTAAGACTGATTTTTGCTAAGGGTGTTTGTTAGCAGCTATGCTGGAGCAAA[T/C]TCTAAGAAACTAGAGGTCCTGGAAATCTGAATAAATCTACAGGTGAAGACTACTCCTTGAACTTGGGAAAACATCAGAATTGCTGTTAATGTACAAATAArs6929257TAAGTGACAGCTCATTTTCAATTTAATTCATTAAAGTATTCCTCTCTACTCCAAAAGAGATAAATAGACTTTGTCAGAGATTTCTGCCTAAGGTGTTAAA[C/A]ATTGCTCATAGTTCAATGTTTAAATAGTTTAAAAAACAGGATCATCATGGCAGATGGGAGGCCGGACTAGATTGCAGTTCCGGACAGAGTAACGTGCGGArs2657300AGGGTCTCTTTTCATACACCATAAAACAGAGACCCAGAGGCAGCTCGCAACTTGCTCAGGTCATGTATTAGTGAGCATCAGAGGCAGGCCAGGACCCCCA[A/G]TTGCACAGGTCTAGGAAGCACCATTTCATCCAAGTCTATGTTGCATGCCAAAGAGTGTCACTGACAGAGAACACAGTGAGACCACTGCTACCGCCCTGGArs7703746GCCATCCAACAAAATTATTATCCGTGCTTGAAAGTTCCCTTTCATTTTTTAACATATGTTTATACATATTATATAGGTTTAAGTAATGATGCATTTACA[G/A]TTTAAAGACTGATGTTTACTTGATAATATATCATACTTACTTGCTATTTTAAAAACTTCATAAATTATAACAGCATGATAAACCATTTAGAGAATCAGTArs7151741TTTGTCTCTCTTCCCCTGCCAAAGAACAGTTGCTCACAATGGGCGTAGCCCCCTTCTGTGCGATGGCAAGGGTGAGGTGAGATAACGTGATCCATTTAAT[T/C]ATTTGGGCACTCAGTGGTTTGATTTTGAAACTTGTGCTGGAACCAGTGAATGTTTTGGGGTTAAACTCCACTGGCAGAGCAGTTAGTGATCCAAGAACTCrs6468296AATCCATCTAATTTTTTCTCTGGTTTCAGTGGCAAATTCAGTTCCCTCCTGAGCCTAGAACCTTGCAGTACATTGTAGATCAGTTGGCTAGATAACTGAA[C/T]TTCCTGCTCATTCCTTTTCACCTCATTCTTGTCAGTTTCCAGTGCCCCATATCTGGATCTGAGCCCCATGGTCTCAGATGCTGAATGGGGCCCTGTCTGArs2846589ATTAAGGGAAAATAAAAATGCTTCTAGATCATTTCAAAAATTCAGAATGAAAGTAGTGATATTGAGTCTCACCTGAAAGCAAAATGTGTATTTTTACAAA[G/T]TATCATTAGTGAAAAAAGAATGATAATGAGAAAGAGAATAATGAGAAAAGAATAATTAGTCCCTAAAATGACAATATTTGGGCACACTTGAGAAGTAATGrs3816551TGGGGTGAGGGATCCCCTTCACTCAGCAGGGAGGGTGTTTCTTTTTCTATAAGTGATTGGGGGGGCATCTCTGGTGGAGATGGGATTCTCTGGTTGTAAA[T/C]TGGGTTCCTTTTGCTTGATGGGGATGGGGGTCTGTGTGTGTAGACTGGGTTTTTTTGTTTGTTTTTGTTTTTTGGTTTTTGGTTTTTTTTTTGAGATGGArs6431221CCAGACTTTGAGCCAGCCCAAACTCCACAGAAAGGAGGGGGCTGGAGAGTGCATTCAATCAGATGATCCATATTCAATCAGTCATGCCTGTGTGATGAAA[C/A]TTCCAAAAAAAGTCTGGACACTGAAGCTCAGTGGAGCCTTCAGATTAGTTAACACACTGGCGTACCAGGATGGTGATGCATCTTGATTCCACAGGGAGAGrs6582294CGGAACCATTCCTCAGAACCACACCAAAGAACTGGCCTGAGCAGGAAGTTACCATGGCCACCACCACCCCCACATTAGCAAAAGGAATGAATCATCCCCA[G/A]TTTGTTTCTTGCTACAGTTCACCTCCACGGTTAAGTCTCACTTCCGCCTCTAACTTACAAAACCATAGTTACACATCTGCAGCTTAGCCACAAGGGAGTCrs3913810TCCTGTGAGAACTCACTATCATGAGAATAGCAAGGGAGAAATCCACCCTTACGACCCAATCACCTTGCACAAGGTCCCTCCTCAAACATTAAGGACAAAA[A/C]TTCACATGAAATGTGGTTGGGGACACAGAGCCAAACTGTATCATCTGTCTAAACAAAAGTACTTTGGGTTTGATTGGTTAAAAAAACACAAAACTTAATTrs7769867ACATTCTTATCATCATCCTGTAGAGGCAAATTTATTCCCAAACATAATCACAGATTACCAAAAATAAAAAAGTATAAGTATTGTCATCCATGGATAGGGA[G/A]TTTATTACATTTGCCTTACAATGACCCAGATAAATGTAATGAGAATGAGAGAGAGGGGACAGAGGATATTATGTCTCCCAAACCTCTGTCTCAGCTACTArs4674824ACTTTGAAAGGCGGGACTCTCTTATGTAGTGGACTTAGAACTGAAGACATGACTTCTTAGTAATGAAACTGAAGGTAAGTACTTGTTTATACAACAAAAT[T/A]AAAAAGTTCTATACAGACTTCTGAATCATACTTTAAAAAAAAATGTGATATTACTGTAACCCTTACCTTCCCCTACCTCCCCAATATCTGCAGTCCATAArs7294836TCTGCACAGGTATCCTGGAACTTAAAATTAAAATATATTAAATTAGAAACAAATAAGGTTTAACTCCCTACCTATCTCTTTGAAAAGCCTTAACCATTAA[T/C]TGAGTCATGGCATTTTTAAATGGACTATTCAGTGGCTGTGGAGATGTGTGCTGTGTTTGCTTGGTTAAGCAGAAAGTAAGTTTTCAAGGATCTCCTGCCTrs6043856TAAATTTAATATACTCAGTCTGGTTCACATATTCTAATAAAATCCAGCAAGCTTTAAAACTTTTATAGGAAATGTGCATTTAAAATCCATGTGATATTCA[G/A]TTTTTACAGGGTGACGTCCTTGCTCAGGGTATTAAGTAGTTTCAGTGATGACGATGACCCAGCCTGGCAGCAAGCTTCTGGGGAAACCTCACAAATAGACrs7741525TTCCACTTTTTTTTTTTTTTAACCATTTAAGCATTTTATTTCCTGATAACCTCTTGGGGTGGAAGGCAGAGTGATATACTGAGACAGGCAGTAGCCTAAT[G/T]TATCTCCTCAGCAGTGACCCCTTCTGAGTGAAGAAAGCAGGTGTGACTGTCTCACTTTCTCACGGAAATAGAAGATTCTCATGTAGCATATGCAAAGACGrs7084321CCAGCCTCTTCTGCCATACTGGGCGCTCACTCCCTGACTCACCATCTCTTGGCCTCACTGGCGGCCAGCGGGCAGGTTTCCCAAGCTAGACCCTTCTCCA[A/G]ATTGCACAGCTGCGTCTTTTCCCCAGGGCAGCTCAGCACCTCGCACGTCCTCAGCTGTGGTGCTTCTGTGGCCCAGGGATCCTGTGTATCCCAAATTCCTrs2723307AAACTGACAAATGAATTTCCATCTTCGATATTGTGTTTTTAATTTCTATAATTTGCATTTGGCTCTTCTTTATAGTTTCAATCTCTGTGCTGAAAATCCT[A/T]ATTTATTGAGGCATGTTTCCCATTTTTTCCTCTTGATCCTTTAACATATTAATTATGATCTCAAAAATGTCCTTGTCTGATCGTTCTAATAGCTGAATCArs4589569GGAAGCAGCCACTTTTCCCAGTCTTGCTGAGACCAAGTAACCCCAAACCCTGGCTCAAAAATACTGTATCAGCAAATACTCCAAGTAGAACCAACCAGAT[A/G]ATTTTCTGGCACTATCGTCTGAATGTATGTGTCTCCTCAAAATGTATATATTGAAATTCTAAACTCTAAGGTGATACTTCTAGGAGGAGGGGCCCTTGGArs3912319ACCTCCACATATGGTTCTCACTTCCCTTTCTTCTTCACTTCTAACTCATTCACCTCAGTAAGGTTTCTTCAACACTGCCCATAGACTCTTTCTTGAGGCA[G/A]TTGTTACTTTCTCTGTTGGATTTCTCTTTTTACTTACATCTGCATGGCCAGTTCTCTCATTTCTCTCACGTGATTATTCAAAAGTCACTCTCTGAATAAGrs2821312AGTAGAGGTTAGACCAGAAGCAAGAAGACTGGAGAGGCATGTTTAGTACCTGCAAGAAGATATTTAATAACTTTCTTAGAGGAAAGATTCAGAATCACCA[G/A]TTACATTGAGTTGAATGAGGAGCATTAGCCGTCAAAAGAATCTGTTTTTCCACTTGTTGCTTGGATAAATGGATAAAAGCCTGCCAAGGACTTCAGTCCArs2820107AACTAATAGAATATCAACCTAGTTAAGGCAAGATTTGCTGTGGGGAAGGGAAGGTCCTGAGCCTCTATTTGGCCCTGAGGCTTATCAACCTGATACTTCA[G/A]TTGGGCTCAGAAAGACTCCTGCCCCCTCCCCATTCCCTCCCTCCCATTGTGAGGGACCAATCTCTCTTATTCTGTGTTTTCTTACCATCTCTTCCACCACrs7002630TTTCCATGTATTCTCACAAAACCTCTCACAGGAATCCACGGATTCACTAGAGGTATGTGAGGAGGAGGAGGATGTTGATGAGGATGAAAACTGACATGCA[T/A]ATTTAAACTTCTACCTCTAGAAAGCACTGGCAAAAAGTAAAGGCACAAGTCAAGAACACGGAAAAAAATCAAGTACTTCCAATGACATTGGCACCAGGACrs7041138CTACATTCATCCTTTCTCCCTTCCCTTCTGCTAAATCGGGTAAAGTATTTCCCTGGGAAATACTTCCCTCGGGCTCCTTCAGGCTTTAAATACCTTCAAA[T/C]TTCTCCCACGTTAAAAAAAAACTAAATATTCACTGAATCCTTACCTGCGGTTGTATCTTATCCTCCACCTTCACAGCCAAACTTCTTAAAAGAATTGGCTrs6019378AAACCCATTCTTCTTGGCTAGAAAATGAGGAAAGACCTGTTAAGTTTCTCAAAGACAGGAACTTCCTGAAATACGACCAGAATGGAAGGAGGGCTTGGAA[C/T]TGGCAGTCCGTCCATCCATCCATCCATCCATCCATCCATCCATCCATCATCCACCTTCCCAGGGTTCAATCATTCACTCGAGTGAATTCTATCATTCACCrs4815732ATTTGTATCTTTCTGTGAGTGCCCACTGGGCCCAAAAATGTCTTTTCTGAGCTTCTATGAAACGTGTGTTTGAAACTGTTCTACGTGAAACATGTTGAA[T/C]TCGGTGAAAGAAAACAGAACATCATCAAGGTTTTTGGCAGAAACCAGGATTTTATCTTTTAGGGCTGGATATTTGGGCAACATAGTGGGACCTCATCTCTrs4488809CTTGTATAGAAATAATACAAGGTACAAGGTGGGGCTGAAAAACACTGGACAAGCTAGCCCGGGGACTGCAGATGCAAGCATCTGCTCTTGAGGCAGTAAA[T/C]TGAAACAGTGATTCTTATTACTAGTTTCTGTTAAAGTTCCTCAAACTCTTTTACACAGAGATATATCTGACTCTTAGATGGCAGCTCTAAAATTTTGAAGrs4420719GCATGGCAGGGTCTTGAAGTAGATTTGTGGGAATTTCCCTGGGACCCTCAGGTCACATGACCATTTATTGGCCCTGCTCTGCAGGGCACCTTAGGTGATG[T/A]ATTGCAAAGGCAGATCAAGGGAAAGCAGATGTTGCCATGGAAATGGCCCTGCCCCCATTCATTATTAAAAGTTGTTGTTATTGTTGTTTTTTGGCAGCATrs6488494GGACACCGAGGAACAAAAAGGTGGCGTGACTTGCCCAAGGATGTTGCAGGTTAGAAGCTGAGCCAGCCGTGGCTATCCATCCTTCAGACACCCAGGCCAA[C/T]TTGTCTGTACCACCTCACATTGCCTCAAACCCTGTCTGCAGTTACTGTCCCATTTCTTTATCATTTGGTCCTGGTATGACAAGGTACAGTGGAAAGAAAArs2522215TGTACCCTTTACCAAAGCTAAACAAATTGCTACTTGCTGCAATCCCAGAGGGGTGGGGGTGGGGGTGCGGCAGGGGCATAGCGCCTGTGCTTAACTGTGA[G/A]TTATTAATGTGAAAGATTTGCATGTGCTTTTTATGATTATAGCTGACTTTGATGTCTTTGTGCCCTGCTCAGGGCTTCAACATACAACTATAATTTATTTrs6142841GTATGAGTGAAGATTCAGGAAACACTGGTTCATGTTGGTTCATGAAATGTTAGTTCCAGATATTCAATAAAAATGTGACGTTTTTAAGAGTTCTAAAAAT[A/T]ACTCGCACTTCTCCACACCGTTAAATGGACTCTCACAGCCTTGAATGAGAGAGCAAAAGTCCCAGCAAGGCCACCCCCTCCAGGTTCCCAAGGAACACCCrs4952502AATTTACTGCTGTTTCTTCATTTTTTTATTACTACATAATTAAAACCTACTGTTAGCTAGCACTGTGGAAAAATGTTGGTGTTATGCAGTATGCATTTCA[G/A]TTGGCTGCACAACCACCATCTCTTCTTCCAGTGACCCCGCTTCTCTGTAGTGATGTGGGTGCATAATTCTCAGACTCCATTAGGTGGCTGTAATCCACGGrs7820949CTCCTTTGGTCAAAAACTTGATATGCCCATTTTGCTTTTCATGAATCTTCAAGACTCGGTGAACTATAGGAATCTTTCTTCCTTCTATCCTTAGAACAGC[G/A]ATTTCTCCCACTCGTATGGGATCTTCAACTCGATTTGTTAGAAAGAGAAGATATCCTCTATGAAATGCAGGTTCCATGCTGCCACTGAGCAACACAATTGrs7076662GTTTTTGTGTCTGGGCCTCTGGGCCAAGCCCTGGACAATATTTTATGAGTTGCTTTCTACTGCTTAGTAACTTCTGTCCTTAAATCCCATCTTGAATGAA[A/G]TTAATGTATTGCTTCATCCGCTTTCCTTGGTGTTTAAATGAGAAGTCTTTGCACCGTCTCTTTGTTTGGTGGTTCTGTCTTATCTTAGCTGGTATTTCTTrs4684986AGGCTAGTATACCACTGGGGTGGGGGAATGGGGAAGTAACCGAGGAGACCCCCATGTGAGGAGGCATGTTTACCATTGGGGTGGAGGTACATGAGGGAAA[T/C]TGAGGGGACCCCCGTGTTAGGAGGCTTGTCTGGCATTAAGAGAACCACGTGGAGAAAACTGAGGGAACCCACATGGGAGGAAACTGATTTACCATTGGGArs6707911TGAAACAGAAAGGAGTACATGATTTGTGGCCTGAGAAATGGCAGTAGAGACAGAATGTGGGCCCAGGTATGTAGCTGCAGTACTTTCTAGGCCATAGAGA[G/A]TTAGTGTCTAGGAAATAAGAAAATAAAAAGGAGCCCATTCTTCTCTAGTGATTCTTTACCACCAAGAGCTAATAAATGTGCATGGTACTGTTCTGACAAGrs4845519ATTATTGAATTTGATCATAGCCATGCAGACTGTTCAACTGAATTATAACCTCTAGGAATATCTTCATTTTAAAAGTGTTCATACTGTAGGCTTGAAGAGA[A/C]TTTAGACATTATCAGTTCTTCATTCACTCTTTCTGTGTCAGCAAGAAGGGGGTTGGTTTATTCAAGGATCCATAGTAAGTTTGTAATGGGATAATGACTArs4420242AGGAGAGCTGGGATGATGTGGGCAAAGGGCGTTCAGGGAAGAGGAAGCAAAGCTAAGGGCCTAGAAATATGAAATAACCTGACAGATCCAGAAACAGGAA[T/G]TGGTGGGAAATATGGCTTATAAAGTGGTCTGGGCCTGCGCGAGAGCTCTCATGGCGTGGGGTGGAATCCACATTCTGTACAATGGGCACGGGGCAGCCCTrs7323716ACAGTCCATTCCAGTTGTAATGGCAGCCCTAGCCTAAGTTCTTATATCTTAGGGACAAAATCTAAACCAGTGGATTTCAAATCCGGCCGCACATCAGAAT[C/T]ATGTGGGGCACTTTAAAAACTACAGAAGTGAAGACCCCATCCAACACCCTCTGAACAGATTTCCGCTAGTCTAGATGATTATTGATTACTGTAGCTTCAGrs6878291GTTTGGATGAGATGTCATTTACTCTAGAGCGTTATCTGTGTAGCTCTGTAATTTCTAATATTTTCACTCTAAAACAGGACAGTAAAAAAACAGGTGAACA[G/A]TTATATAACATGAACCAAGATTCTATGAAGAGTTTTAAGCTTTATGAGGACAAGGACATCTTGCTTACAATTATCTTCAAATGCCTACCACAGTCTCTGCrs6897414CTTTGAGGTTTGTTTAAATGTTGCTTTCCTTTCTGGGTTTCTCCAGGTAGCTCCTCATCTTCAGTGTTCTCATTGCACTTTGTATTCTCTTAGCTTTTAA[T/C]TCTCAGGGACAAGGACTGACCTGGTTCAACTCTGTATTCTTAGGGCCTAGCAAAGTGCTTAACACATAGAAGACATTTGCTGACTTACAGTTGAATAGAArs4452041CTTGTTGAAAAGTCCTTAGGAGACTCAGGGACATGAGAATAATGTGTCCAGTGACCATAAGGCTGTCTTTAAAAGAAGAACATCCTGTCAGTGAGGAAAA[T/C]TGCACCTTTGTCACTTGCTTTGCGTCAACTTCAAGGCCCGACTTAACTATTTCCTCAGCATTCATTTAGCACTATTTATTAAGCACCTGTTTTAGGTTCTrs7144509TTTTTTTTTAATGAGAGTTGATGGTATACCATCTTGGAGTGTTGTGATGCATGCCAAAATCAATGAGTTCATCTTCAACATATTGAAGTTCTTACTTTCA[G/A]TTTGGTCTTAGGAGTGCCAGTTGCTTAATCAGGATAAAGTACAGTGACTTAATTACAATGCCAAAACAGATAGGAAAATTAAATTCAAAATCTTAACTCTrs7845628GACCTAAAAATTTATAGTATTGATTTATTTTTTATTACATGGAAGATGCACCACTTTCTGGACATAAATAAATACATAAATAAAATATAAGCCATTTAAT[G/T]CCTCCTCTTTCTTCACATTTTTCCCAACCTCTCCTAATCTTTTGTTCTCCCACCTCTGCCTGTCTTATCTATGTCCTTGCTTTACCTCTTTCTTTCCTTArs2903113AGGGATGCCTGAGGAGCAGACAGCATGAAGGAGGGCAGAACCGTCGTGGGATCCTGGTGACACTGTTCGCATCTGCATCTGGCCATCCCAAGCCACAAAT[C/T]CTGGACTTTCCCATTGATGTAAACCAATAAAATCTCCTTGCCTGAGCTACAGGAGGTTCTGTTTCTTTCAAATGCAGACAAAATATTTTGACAAATTACCrs7831906CAGCCCTCTTTCAAATATACTCTAGTCTAAACCCTTGATTTTAACCGTGTTATAGGGTTAAGTCTTTTCTGCTTCTGTCAAAAGCCAGGCTAAGGCAAAT[C/T]CATCAGGAAAAACAAGACTGGAAAACAAATGTAAACTTTATACTCTTTGAACCTCTTTAAACTTTATCCCTGTATTAAATTTGATCACAAGAAAAGCTCArs6595267CACTGGAAGGAATAAATTCCAGACACAGTGGCAGCTGGCACTGTGCTGCTTACAGATCAAAGACCTACAGGATTACAAGTAAGGTTGGGTGGTGCCTTTG[A/C]ATTCTCCAGGTGGTCTTCTGTGTCAATGTGGAGGTTCCATGAATAGGAATGTAAAGGTCCATGGCAGAGGTGTGGATCCCTGGGCATCTAACTATCACTCrs4783152AGAAAAAAGTCTTCAAGCCTATGGTTATTATAAATCCTACACGCTCCTGAATTCAGTCATGCCAAATGGAACCAGAACCATGTTTTTAACCCTTTTAAAA[T/C]TGTGGTAAAATAGTCTGGGCACGTTGGCTCACGCCTGTAATCCCAACACTTTGGGAGGCCAAGGCGCGTGGATCATTTGAGGTCAGGAGTTCGAGACCAGrs7094883GGCAAAGGAAAGGGATGGATGCCCACAGCTCCTGTCGTGCAGCTGGAGCCTTGCAAAGCAACTTCATAAAGCCTGTGGACTGTTAACCTGCTGACTTCAA[T/C]TGAAAACTCTTCAGATGTTAAAAACAAACCTGTTTTGGGCTTAATGTCCAGCAAATGTCATGTTTTGCTAAAAGACAGCATGGCAGGCAGACCCCGGGGTrs2804649AAGCTCCCAACCTTTCAGCAGCTTCTACACACCCAGCTCCTGCCACCCAGTGGCCTCTTTAGGCCAAGCTCATGCTTCACAAGGGTCTTTCCAGGCCCAA[T/C]TTTTGTCTCATGGCAACCTTCCCTGGCCAGATTCCTGCCTGTCTCCCAGCAGCCTAGACAGGCCCAGGTCTTGCCTCACACTGGCCTCTCTACATCCAGCrs6569474CCATAGAGCCCCACTAAATAATATAACAGCTGGAAGGGATTTATTCATCTCTGGACACTAAGGAGTTAGGGCACAGTAGTTCAGTTACCTGGTTATATAA[A/T]TCTGGGAACCTATACAATGATTAAAATGGAAATGAGACCTCCAGTTACTGCAATGAAGGTAAATGGTTTTCCAGGGGAATTACACTTGGACTCAAAATCArs4869315TCAAAATCTCCCCACTGGCGCATTTTAGGTGTTTTGATCATGAGTCACCAGGAGCTCTAAAGCACTTAACTGAGTCTGGGGATTTCTAATCTTTTCTGCCA[G/A]TTGTTTGTAGGGAAGTGCTCTGTGAGCTCTACCTCTGAGGCTCCATGCTCCCTCTGGCCCTCCCTTTAATAGCTTCTCTTCCACGGAGATGCAGTCAAGTrs2937415TCGAGGTAGGAGGTTGGTGTTTGATGGATAACTCTACTGTATAAAGTTAAATTTGACTGGTTTTCTATTTCTGAATCATGGAAGTGATGAGAGGAACTAA[C/T]TGATTTATCTGAAGTCTGGATATGTAATAAAGTCTTCATGAACTGCAGTTGAATGTGGCTGCATTGTTACTAATGTACAGAATTTTTTCCATATTGGCTTrs7737946TTCTCTTCTCTCATGCAATCATTAATTCAGCGTTGCTCCAACGTCAATGAAGCCTAGTAAAGCTTCATCATGCTTCACGTCAGACTACACTGAGCTACCACA[T/A]TTACATGGGATTAAAGAAAACTATTTGGGGCTGGACCCAGTGGCTCATGCCTATAATCCCAGCACTTTGGGAAACTGAGGTGGGTGGATCACGAGGTCAGrs4311632TTTACCCCAAGTCTTCATTTTTTTCTCCCAAATTTATTCACCCAAATTCTTTATGGTTTATTGGAAATGAAGTCAATATTTAAAGTGCTACATCTATGAA[T/C]TCAAAGTTCACATAAAATCTACATCAAAGACTGGAAGTAAGTAGGATCCTTTAGTGGTCTAGCTCATTTGCTTCTCCAAAAAGCATAATTTTTCATGAGArs7356482GTACGTTTTGCACACATATCCATATTATTTCAGTGTATCCCTCAATTCTGAACATCAGCAATATAATGCCGCATGAAACAATCCTTTATCTCTCTCTAATAC[A/T]ATTGTCCCTGTGAACAGTGATCCACAGTATATATGTGTTCTGTTCTATCCTTAGCCCGATGACCTCTGTCTCCCCAGGAGCAGCCTTCTCGTCATTGGCArs4928169TTGCTAACATAATTTCTTTATCTTCTTTTAATTTCCTTAACATAGCTCTCCTAAAACTATGGGTAGTACATGGGGTTTTGGGGGGGTCAGGTAAGGAATA[A/T]TTACATTTTGTGTGAATATTCAAATGATGCTTGGTAAAAATCTTTGATAACTTCTTAAGTGATTATTTCTCTCCTAAAATTCTTTGAATATAGGCATGTGrs4673821AACGTGCCCAGCACCTTGGCTCATCATACCACCGTCTGGCTTACCTGCTGCAGGTCACTGAACTCTGGAGTAGATTGACATCAGATAGCCTCTTGTGAA[T/C]ATCCCTAAAATGATGGGTTTCCTTTGAAAACTGCTAACTCTTCATACATTTCTACATATACTTTACTGCATTCTTCTGTGATTGAAATTTGCTTCTTAATrs7588807AAAAAAAAAAAAAAAAAGAATATAACAACCATCCATGTAGACACCATTCAGGATACTGTTTCACACTATCTGGTGAGTAAGCATTGAAGTCTAATACAAA[G/T]TCTTAGTGCTGGTGAAGTGGAGAGAGGGCCTCTTACACACTGCTGATGGGCATGTTAATTGGTAAAACCACTTCAGAGAGAAGTTGGCAATGTCTTCTATrs7679285TCCAGAATGCTATACAGTTGGAATCATACATACTGTGTGATTCCAGATAAACTTTGAGATAAACTTATTTAGATTAGTGATATGCATTTATGTTTCATCA[G/A]TTTTTAAAATACGATGATAGCTCATTTCTTTAGCAGTGAATAGCCTTCCATTGTCTGGACATACCATATTTTATTTATCTGTTTAACTACTGAAGGACATrs7688917GAATGGCAGTAAATATTCCTTGGGTTTCACTGAAGTCTGCTCTAAATGCAGTTGAGTTTAATGATCAAGAGCTTGGACTCTAGCATCAAATGTGAGTTCA[G/A]ATTATTTCTCCATTACCTCCTAGTTGACTAACACCTACTTTCTGACTATAACAACAAATGTTACTGATAACTATTTCTAGGAAATTCATTAATAACATATrs4533845ATTCATCGCCACGGGGTGGTTCTTTTCCACCCAACAGCCTAACTCTGCTGCTGCTCCCAAGGGCAGAACAAGGACAGATAGGTGGATGTTTCAGGGAAGTAAA[G/T]TTCAGCTAAATATAAGGCAGAACTTTTACTAGACTTTTTAGAAGAGTCAAAAAAGGTATTGTCTTAAAAATGGTGGCTTCTGTGTCACTGGTTTTTAAGCrs7205009ATGTAGAGTTATTCCTGCAGGGCTGTGATTATAGGCAAATCATAGTGTGTATACCTTCTACCTTTTTTAGTGTATCTCCCACTCTTGTACCCCAGAAAAA[C/T]TAGTCTTGAGTATCCTTCCAGAAATGTTGTATTCCAGTCTTGTGTATCCTTCCAGAAGTATTATAAATATTATATTATTATCCTTCCAGAAAAATCAGTCrs7604667TCTTTTTTCTTCAAGGGCACCACAAGTCTTGGGTGCTGTGGAGAATTGCTGATTATTTTTTTCTCCCAGACATATAATACCTTGGTCCCTTATTGTTCAA[T/C]TGATGTAGCCTTTTCCAATAGGCTTCTTCAGTACACTCTATGAAGGAAGCAGACGTAAGAGTCTATGTACGAGTGAAACTTACCCTAGGATACCCACTTCrs4442368AATGTCCAGGGCCAGAGGAAGCAGGACAGCACAAGATTTTATCTTGCTAATCAGAATGGCAGAAAATATCTCTTCTCTGTAGACAGAAGACAAGGTGGCA[A/G]TTCTAAAGAAAAGAGGGTGTTCTAATAATCTTTACATGTACTTTAGACCACACAAGGAGGTAATATATCATTATGTGCTTTTGGCTTCTCAGATATTCTArs6575809TAGAGTCCCACACACTTACTTGTACTAAACATTAACCTGCATGTCCAGTCCTACCCAGTACCTGAGTCAACCTTGGAAAGATAAGAGAGATATCAGAAAT[T/G]TCACCCTCACCAGCAAAGGGGGTGGAGGGAAGACTGTTGGGGGAGCTATTAGAGAGCATCTAGAACACCTTGGCTTATCATCTGATTCACCAAAGGTAAGrs6807437TCATAACTGTCCCCCACAAAAAATAATGACAATTGTGCACCAACCTCCTAACTTATTATTATTCTTTGCTGTGGTTCATGAAATCACAAAGTCTTAGAAT[T/C]ATTGCATTAAGGTACTGCCACACTTAGTCCATTCAGAATGCCTAGACTCCCATACTGGTGCTATCATTGGCCTCAGAAGGCATATAAAATGAAACTCAGCrs3902595CAAGGCCCAGGAACAGGATGTAAGAAGGGAGGAAGAAGAAAAGCCAAAGGGAATCCTCTTCCTGTGTCTTTAGGAAGATCCTGGAAGGTGCTGAAGCTAA[T/A]TACTTGGTGTTGATCTGAAGTTAGACACTTAGCTAGGGAGACTTGTTAGTATCTTTTTCTTAAGAAACCATGTGCTGAGCTAGAACTAGTACTGTAGTACrs7763815AATGAAAAAAGGCAATGAGAAGTAAAAATGGAAAAGTACCAGTTCAAGCATGGCAGCCAAACATCCAAAGACTATCATTTGATAAAAGATTTACCTGAAT[T/C]AACAGAGCTTTCTTGACATTGATTAGGGTGGTGAAAATGACTGTGGAGGAAAAATTAAGTAGATGATTCTATTTAGGGAAGAAAAGGTGGGAGAAGTGGCrs3010003AAGACTTTGTCTATCACAGCTCTTTCAAAGTGCAATGTTGGTGAAGGATGTTAACTGCAAGCTAAGAAACACACTGGGTGATTTATGCAGAAAAAGAATA[C/A]ATTGAAAGCACCCTAGGTATATGGTACGTTGTTTATAAAGAAGGCTGTGATAATTTCTCACATCCCTTGTGGACATGCTCCTTTACCATGTGATCTTCCArs3902451TTGAAATATGTTTTCCAATTTGTTTGCTTTCTTCCCCTCCCTCTCAGGGATGCCGAGGATTCATAGATTTGGTGTCTTTACCTCTTTACATAATCCCACA[A/G]TTCTCAAAGGTTTTGTTCATTCCTTTTATTCTTTTTTCTTTGACTGTCTTATTTCACAGAACCAGTCTTCAAGCTCTGAGATTCTTTCCTCAGCTTGGTTrs4683161TCACTGGCCCTCAAAGCTTTTGCTCAGCATCTACTTATGGGAAAATGCAAGCTACAATGGTTGAACTTTCAGCTTCCATCAACTTGAGACATGTTCCAGA[G/A]TTTAAAATATTCTTCACTTGTATTACCCCTGTCCACAGGCAATGAATCTCCTGCTGGCCACGTGGCTAAGAGACTTGCACAGTCCTAGGATCCTTGATAArs7691446CAAACTTTAACACCCTTCTCTGAACAATTGATAGAAAAACTAGATAGAAAGTCTGCAAGAATATAGGATGCAACACCACCATCACAAAAAGAGGCTCTAA[T/C]TGATATTTACAAAACACTCATTGACATTTACAAAATATTACAAACAACATGCAAACATATATTCTTTTCAAGTGCTTATGAAACACATACAAGATAGACCrs4974594GATGGAAGCCCTTTGTCAGCTGTGCTGTGAAGGCCTTCTCCCAGTCCATGGCTCGTGTTCTTAACTTTCTTACATTGTTTTCTGAAGAGCAGAAGTTTTA[A/G]TTTTGATAAGGTTCAGAGGATGGATTTTTCTTTTACAGTTGGTACATTTTGTATCCTTCTAAGAAATTGTTGCCTGTCTCAAGGATGCAAAGATTTTCTCrs6139756GACAACTGTGACCCGAATTGTTGCAACAAATTATGATGTAATAATCCTGAGATCTAGCCACTTACAGAACAGAAGGGAAGACAGCAGGCTGCCTTGAAAT[C/T]GAGCTGGCAGATGTGGGTCATTGGGGGATGGGTATAATATGAGAGAAGCTCCTTGTGGCTCAGCTCAGAAAGGTTTGCATGTGTAATACCAATTATTCCGrs2889515TCAGGAGAAAATATTCGGAATGAATAGAATAGAGAGACTAAACGAATGGAATATAGAGAAAAGAGAGAATAACAGAGTACACAATGAGAAATGTGAAATATTTGATAAT[G/T]GAAATCTTAGAAGAAAAGGAGTGAGAAAACAGGGGAAAATAAATATTTAAAAGAGAGTGGCTAAGAATTTTCAAAAACCTGATGAAAGACACAAAGCCTCrs6494229GCTAGCTGTGGCTCAATATCTATTATCCCCCTTCTTCCATAGCCAGAATAAAGACCATATTTTCCATCCTTTCTTGCAGCTTAGTGCAACTTGGTGACAAA[A/G]TTCTAGAAAATGAGATGTAACTACAAATGAAGCACACAACTTTCAGGTGGGGCCCTAGAATGAGGCTGACAATCTGCACTCCCAAGCCTATCCCACACCArs4678766GTGCTTTCTATCTTAATGTATAATTTATAGAAAAACTAACTCCCTTTAGGTTTTGGCCAACTTGCTCATGCCGACAAAACTTCCTTTAAAATACCA[G/A]TTTTTCACAAATCTACTTTTTCTTTGGTTTTATTCTACCATTCTTTTAACTTAGGACAATCCTTAAAATCTCTAAATGAGACTGAATTACTTTCCCTTTArs2984523TTATCTCTAATTCTTACAATAATTCTTTGAAGCAGTGATGATCATCCTCTTTTGCAGAGGCAGAGCTGAGGCACAAGGAGATAAGTAACATGTTTAAAAT[T/C]GTATAGTTGCTATCTGAGAATGAAGTCAAACCCAGGTCAGTCTGACTTCCAAAGTCGAATTCTTTCCAATATAGTAAGTGCCTTTCCTAAACCATTGACArs4130306GGCATGTAAACTCTACCACAAAAAGATAAATATCATTTCCAGCAGACAAATATATGAAAACAGGTATAAACTGATGGCTCGTACTACCCAGTGGAATAAA[C/T]TCTTCTGCAATAGAATGAATATGTTCTTCTATAAAGGAAAAGAGTCACATCATAGGGAAAAGAGCTTATTTGGTGAGCACATTTAAAGCTGAATGCGTATrs4889072AGGAGCAAGATAACACAGGGCTTTCTGTTACCTTGCTTAAGCGGTGGGAGAATACACAGTAAGTTCCCTGAGGGCAGGGACTATGCATATTCTGTTA[G/A]TTTCTCCATCCTCCAGATCTCATATACTTCCTGGAACATATTAAATGCTTAGTAAATATGTGATAAGTGAACATGAGTGACTGGGAAGAAAGGGGCTTAGrs6005754CCACAATGAGAAGTATAAATCTACGAGAATACGAGAATACACTTCAAACTGGTAACAGTGGTTACTTCTGGAAAAGACAGTTTTTAAATCTTTTACATCAATAACTAA[T/C]TCATACATTACTTATGTAATGATAAAACTATAACAATAAAAAAACAAGTAGTATGGGAATACAGATGGCTGAGCAAATAACACTGTTCCCAGGGTTGCTGrs2734574ATCCAGATAGCGAGCTGGCTAGCAGCTGTCCACTCTCCAGCAATCCTGCCTTCTGGGGCATGGTTTTCTAAGGACCTTCCTGTTCCTAGATGATCAAAAT[T/A;A/T]GGGACCAGCCACTCCCTTCTGAGCCACTCCTGCCTCTGGGCCTGTGGCTATGTCACAGTCCAGTCACAACAGGACATCCCTTCAGAACACCCTGCAGGAArs2734574ATCCAGATAGCGAGCTGGCTAGCAGCTGTCCACTCTCCAGCAATCCTGCCTTCTGGGGCATGGTTTTCTAAGGACCTTCCTGTTCCTAGATGATCAAAAT[T/A;A/T]GGGACCAGCCACTCCCTTCTGAGCCACTCCTGCCTCTGGGCCTGTGGCTATGTCACAGTCCAGTCACAACAGGACATCCCTTCAGAACACCCTGCAGGAArs7725509CGTAACTTTTCCTGCACAGCCTTAGTGTCTTATGCAGAAGAAACATTCGGTAATGCCATTCATTGCTCTACACTTTTCTAGCATCTGATTGTTTAGAAAA[G/T]TATTGCAAGCTCGGTGCAGTGGCTCTCAACTGTAATCCCAGCAGTTTTGCAGGCTGAAGCAAGAGGACTGTTTAAGCCCAGGAAATCGAGGTTGCAATGArs6592545AAATTTAATAAAGAACAAGTTAGGGATCCGATGATTTGACAGTGGATAAGGAAAAGAGAAGTATCTATCTAGGTTGAGACTCAGGTGGCTGGACTGGGGA[A/C]TTTACGATATGCAACAAGTTCAGAAAAGCTTTCATGTTGCTTAAACCTTTAGGCTTGAGAAATAAATATTTATCAGTTGAGATAATTAACAGATCCTGCCrs2676403CTGGGGGTCTCTTATAGATTCAGTCACCATCATTATGTAAACTGTTGAGGCCTTGGATAATGGTTCACTAATAGATAACTATTAGTTGCCTAAGACATTT[A/G]ATTTTTCATATTTTAAGATTATGATTTTCAGCACAGGTTAAAGTATGTGTGCTTTGGGGATATATGTAATGGAGAACAGAAAGAATCCACAACTCCTTTTrs2792780TCACTAACAAACTGCCTCCCCACCCTTCTTCCGCCCCTGCTCAATGCCCTGCACTTCCAGCTGCTGCTTTCTCTGCTTATGTAACAGCTTCCCAATGGCA[C/A]TTTCAGCCAGGATGGGCCTCAAATGACTTTCTGCACTAAATCCCAGACCTTTGTGTAATGCAGTCATTTTGCAACCAGCCTCCCCAAGCTTGCCAGAGCArs9599645AGAAATAAATGACTTTGGTCATGATTGGGTTTCCTTACTTGTCAAAGTGAAAAAAAAATAGACAGATAATAATGTATTAAAGATGAGCCCACAGGCAAAA[G/A]ATTAGTCTGATTTGTATGTCCCTTCTCTCTGATGTCTTTTTAAGGCATTTGTAAAATGTTTTTAAAGAGGACAAGAAAACGGTAGCATTTTTGACAGATCrs10898954TGCTTTAAAAGTGAAATGTTTATGGTACTATGAGTGCAACAGAAAAGGCAGGGGTCAAAAGAGATCTGGGGTCTAGACCCAAGTCTTCTATCAAGGGAAT[C/T]GGCACTTAGAGAACACATACCAAGGACCAGGTGCTAGATTTCAAAATCTTTCTCTATTGCCCGCCTGGGCAGTGTCACCTGTAACTTTGAACTCCTGGGCrs9652080GGAGATTGGGCATTTCACAAATATCTGCAGAATAATTGTATCCATAAGCTATATACAAATATCTACAGATAACAGTTTAAAGTCATATTCACTTTTACTG[A/C]ATTAGCTTTTGGCAACACATTTTGTTTTTTATTTTTCTGTTTCTATAGTCACCAAACTAAATTCTTACCTATTATCTGGTTTCCCAAATAAGCCTACCTArs9352730GCCTCTTCTGTGCATTTCTAGGCCTGTGGTAGTAACTAATGATGCTTTAAAAAATAGGTCACTAGTGTATATTTTGTAAAAAGGGATAGTTGTAGTATGA[A/C]TTGCAAGTCTTGGAGGTATTGTGTGTTGGGAGTCATACTAAGAAAGGAGGAAATTCTGTTATACAGTCATGTGCCATATAAAGGCAATTCTGGCAATGGTrs12818834ACAATGTTCCAAATAAATCAGCAGATTACAAATGAATAATTTTAAGACGGGATGAGATGATGTTGAAGATAAAGTGCACAATGGCAGACCATCCACATCA[A/G]TTTACAAAGAAAAGATCTTGTCCACGACTTAGGTGAAGAGAGCCAACTATTAATAGCAGAAATAAAAGCCAACATTATAGACATCTCAACTTGTTCAGCTrs11835780CAAGACTACCATCCGCGGACTCACGGAATGCCGTATTCACTATCATGGTATTCCACACAGCATTGCCTCTGACCAAGGCACTCACTTTATGGCCAAAGAA[G/T]TGTAGCAGTGGGCTCATGCTCATGGGATTCACTGGTCTTACCATGTTCCCCATCATCCTGAAGCAGCTGGATTGATAAAATGGTGCAATGGCCTTTTGAArs11105611TGTTTTTTCTTAAAATTGCCTTCAGTTTCCTTGGAAATGGAGGAATTTGTGGAGTATTTTAGTTATTCCCTTTCTGGCTGTGGCAACAGAAGGGCAGATC[C/A]ATTTAACTATTTATCTGCCCTCTCAACATATCTTCTAGTTATATTTGTTTTGTAGGCTTCAAACCTGTGAAGAGCCTTGACTGAGGGTTCTCATTTCTCCrs12450474AAATGGATTTACACAAAGTAAACATTAACTTTGGTAGATTTCAATGTAGAATAGTTCATAACAAGCATATTTGCCCTTCTGCTCAACTACCAAGTTAAGA[C/A]TTTTTCAAGTATTTTAACTGAGATTTTATTATGTTGACATTTGTTTCTCATTCCACATCGTCTTTGGCCAAGCGCCAGCACTTACAAGTCTCTGATTAACrs9594249TGTTTCTTCTTGTCTCATTCATTTTACTATTTCTTATACCCATCACAGGGTCTGTACATTGCAGGCATTCAGTACTTTTTTTTTTAAATGAATGAGGCCA[A/G]TTCAGGTTCTAAACTTTTGAGTTTCTTCTCCATATTTCTTTTTGCTTTATTACTGCAATAAATTATTTCTTAAATTCTGTTTAATCAGAAGATTTTAAGArs9285190TATTAAAAGAAAAACTGTTGAGGCAAAAAGAAACAAAACATTTCACCTTTTTCCCTGTAGAAGCCAGAGTGTGCTTCTCACAAAAGCCTGTGCAACCTCC[G/A]ATTTTATTCAAGAGCTAAAGAAACTAGCAGTCTCCAAGGCTCCAAGATTTAATTTCCATTGCATAGGATGCCCCTCACATCAGAATTAATCAGTTTTCATrs17170027AGGAAGAACTCGGGGTGTGACCAGGATTTTCAAAAGCGGGGTCAGAGAGGAACTGATGGAAGAAAAGCTTATAGCTAGAATAGAATAGGAACTCAGAAAA[T/C]TGGTGTTGTCTTGGGCTTATTTTCTTTGGCATCTTGCTCACAAAAGGATAGAATGATCAAAGGATGGATGTCACCAAGAGCTAAGCCTAGGCCATTCCTGrs7899028AATTAGAACCACATATCTTAACTAGAACTACCTAGAACTAAAAGTACTTGTAAAAATATGGCATAGGGACCCCGTGAATCAGCAGGCTTAGTATTGGAAA[G/T]TATAAACGCTCCAGAAATGGGGGCAGGGCATGTGACTGTGATTTGTGGCCAGGATTGAGAACACTGGCCTCCGTGAGCCAGGATGAAAAGCAGCCTCCTTrs11079666TTTAAAAACCAAGTAAACCCCTCTCATTGCACCCCCTGCTACTTCAGAGGAACACCTCCATTCTGATGGAAGGAACTCGTACTTGGGTCCTGGAACCCTA[T/A]TTGGGACCCAAACCTCTCCCACTTGTGTGGCCTGACGTGCCTGAGTGCTGTTTGTGTCCTTTTTATTATGTTGAAAACTTTGTTATTCCAAAGAAACATCrs12034424CTAAGCACTCTACACACTTGAGCAGCTTTTATTGAAAGAACTTTGCCTTTGAACAGAGGGTTTAACAGCACATTATTTCAGATATGTTCAGTCAATGAAT[T/A]TCAGATTCTTTCTTGAGTAGCAAGATATATGAATAGAACTGAGTAAGGTTTCTACTTTTTAAAGAGTGCTGCAATGAACACTCATGCACATGTATCCTGArs10276221TATCAAAAGATGAGTGGATAATGAAAATTTACTATATAGACACAAGAAAAAGAAGGAAATGTTGTCATTTGCGGCAAAGAGGGAGCCAGAAGGATATAAT[T/G]TTAAGTGAAACAAATCAGGCACAAAAAGATGAATATAGCATGTTCTCTTTCATGTGTGGGAGCTAAAAATGTTGAGCTTATATAACCACAAAATTGTGGTrs9886292TATCTCCCTATCAAGCCCTACCATTTCCTCGTTCTCATCATACCCATTATCCCTCAAGGGCCATAGAAACACCTCCCCTTGTAGGACCTAACACTTCTCA[G/A]TTCTTCCCAGGGAAGCAGATCCTGAAAGCCTTTTGGAGGTTTTGTGTCATGGTTATACAGGAAAGAGTATTTAGATTACAAAGTTACACATTGGCAGGGTrs9630712TGCCTATCATAAGCCTAGAGAACTTGGGAGTTAGTGAAATATAACATTCATGTTAATCAACCTTTTAGACATGGTTGTGTTGTGAAGTAAAAGCTGGAAT[C/T]CAGTATTCTCAGTTCTGTATATCATTATCACCAGCGGTGCTTTAAAGAGAAAATTATAGGTCTCTCTACATCTATCATACACCTCCTCAGATTCAATGGGrs10851704TTCTAAGCTCAATAAAGTGCCATTATCCTGTCGGTTATAAAAGAATGGTTTGGAAGATCCTTCACAGCCCACCACTCTCACACAAAGTTTGCCTGACAAA[C/T]TTTCTGGCCAAAATGGAAGGCACTAAAAATATAGAAGTTATTATCAGTCTTAAGACAATACCGTTATATAATAAATAAGACATTACCTAATTAAATTTTCrs10034384TTTGTGGCTGTGCAACAATGGGCAAGTTATTGAATCTTCTTTTTCATCACTTGTGATATGAAGAAAACATTATATCTACTTCCAAAGATTGTTGGGAAGA[A/G]TTAACAAGCTATGCACTTTCATTGTAAAAGTGCCTGGATCAAAGGACTCACTCAATACGTGCTAATAGCTATTTTTTAATTTGCACGTAAGAAGACTGAGrs12442455GTGAGGACAAGGGCCAGTGTGCAAATATTTGCAAAGCAGGAAGACCAAAGAACCTGGCTCATGGAGACATCATTAAGTCAGTGATTATCCATTGGAAT[T/C]GACTTACCTCTGGGCTATGCGTGACATATGAAACTCATTATTATGGAAAACACTTTTTGCTAGGTTTTGTGTACTTGCAGGTAAAACACTCTAATGATTTrs12439908ATGGAGGCAGATGCAGTTTCTCCTGGGCAAAGACTGAACGAATGCAGCAATTAAGGAGACCCCAACTCAGATTGGCCTGATGCCGGGTCCTGGTGCCCAA[T/C]TGTGTCCTAGCTCCAGGCTGTGTCACCCCGAGGGCCTGAGACCCATGCCAACAAGCCAATTTCCCTCACCTGTAAATGGGAATGCCATTACCTGTCCCArs11221268CATGCCACCACACCTGGCTAATTTTTGTATTATTAGTAGAGATGGGGTTTAGCCATGTTGGCCAGGCTAGTCTCGAACTCCTGACCTCAAACAATCCAAT[C/T]GCCTCAGCCTTTCAAAGTGCTGGGATTACAGGCATGAGCCACCATGCACCACCTAGTTGATTTTTGTATTAAAAATGCTTATTGTCCAGTTACATGCATTrs9515625AAATGAGAATTGGAATCTCCCATACGCTGAAAAGAAGTCTGAGACCAAGAGGTGCCAGCTAATCTAACACCACACCGTGATTTACTAATAAGTATCAAAT[T/A]TTTAAACCTTTCCTTTGTGGCCTCATGCTCCATGAACTTTTACCTATAATAAAGTTATTTTTTCAAATAATATATTTCTATGTACTCTAGGGGATACATArs9322744CTCTGCTGGATAACAAGTGTCAGCTGCAAAAAGACCCATGTCTGTCATACTGTAAACACTCAAAATAAAATAAAAAGCATCATTAAAGTATTTAGCCAAT[C/T]TCTTTGCACATCAAAAGTGCTCCATATATTTTAGTTCTGAGTTTACTTATGCTCCAGGTATAAAATTATCATCATTTGAACTGAAACTTTATGATGAATTrs9864594TGATCTCAGCTCACTGCAACCTCCGCCTCCTGGGCTCAAGTGATTTTCCAGCTATTCTCCTGAATAGCTGGGATTACAGGCGTGCCACCAGATCCAGGTA[G/A]TTTTTAGTAGAGATGGGGTTTTGACATGTTGGCCAGCCTGGTTTCACTCCTGACCTCAGGTGATCCACCCGCTGGGATTACAGGTTATCTTTTTTTTTTTrs9356029TCTTTCATATTATTCAAATCTAGAGCAGCTGTTCCTCCTCTAGGTACCCACAGCATCCTGGGCTTTCCTTTGTCATAGCATTTGTCACACCTCTTCAAAT[C/T]TGTTTCTTTATCTCTCTTGTCCACTAGACTCTTGCAGGCCGTATGATACTCTTATCTGCATGCCCAGTGCCTAGCATGGTACCCAGCAAATTGTAGGCAArs10740169TGGGTTGAAAGGACATCTAACTATCTTTAGTGTTTTGTGCCACCCCGTCCTGCTTTCCTCTCTTCCTTACAGAGCACTTGGACAAGAATCCTCATATCAA[G/A]TTTCAGTTCTTAGAATCTAACGTAAGATACTTTCAATCATTATTTCCCTGAAAGAATTTAAGCATTTTCAAAGCCCTTTTTAATTAAAAATAAAAATGTCrs10964719ATTAAATATCTTCTCACTTTCAGCCTGTGTGTGTCCTTAAATCTAAAGTGAGTCTTTTGTAGACGTTATTATAGTGGGATCTTGTTTGGGTTTTCGGAAT[C/T]CACTGTATGTCTTTGATTGAGCAGTTTAATCCATTTACATTGAAAGTACTTAGTGGTAGGAAAGGACTTACTATTGCCATTTTGTTAATTGCTTTTGTCTrs10893402ATGAAATTTCTCACAGTATTCTTTATTTCCACTCTAAAATTACGGAGAGGTAATGAGTATAATACTCAATGTATTCATTCATAGTAGGCAATCAAGCAAT[T/C]GGTTTTCATTTACTTGGTTTGGAAAAGCTATAAAAACCTTTCTTTGTAATCATGGACTAATAATTACAAAAATTGTTTTGTCTCTGTTTCTATACAATACrs10956363TTTCTCTGGTAAGAGCAAGGATACTAAAACATGTTTGAGTGCTGATGAAATTGATCCCATAGAGAGAAAAATGTTGAGAGTACTGGGGAAAAGGGGGATA[G/A]TTGTAAGAATGAGGTATTTTAAAGTGTTAGAAGAATGAGATCCAAAGAGCAAGAACTGGCTTGTCTTAGAGAGGAGTAGAGACAGATCTTCAATTATCATrs11771935AGAGACAGAGTAACGTGTTAAATGATGCTGCAAGGATGCAATCAGCACCTCTGCAAGCCCACAGGACAAACACAAGTGTACAAAACAAGTACCAGCAGTA[T/A]TTTAAAACGACGGAGTGGAATCCACAAGAAACATAAGACACTTGGTATATAAACCTTATTTGGATCTCATTCAAGCTAGCAAACTGTAAGAACAGATATCrs10901705GAATTCACTTTTATAAGATACCCTTACCACACATAAAGCAGAATAATTTTATCTGAAGGTAGACCTGGATGATATTGTAAACTCTGAGAGCAACCACTAA[T/C]TTTTTTTAAAGGTGTGTAATGATATCCTGAGAGATTAGATAAAATAGAACCATATAAAATCTTCAAGTAAAATCAGAAAAGGCAGAAAAGAAACCCCTGGrs9989393TTATTATTATCAGAAACAATTTTTGTACTATGCTTTATATTATAATAGGTGCCCAAACATGTTATGCTATTTGTCCAAAAACACTCACCAGACAAAATAA[T/A]TCTTCTTAGTAGTCCCAGAGGCGTTATGCTTCAGTTTGTTTTTCTCCCTCTTTGCTCCCTGCACTTCATCAGCAAGTTTGTTCATTCTGCTTCTGATTCArs10860857TTCCTCCCATGACATGTGGAGATTATGAGAACTATAATTCAAGATGAGTTTTGGGTGGGGATACAGCCAAACCATGTCAGTACCACTGATGATTTTAAAT[G/T]GACTTTGTCGCTTGCCTTGGTGGTTCATAGAAGAGTGTCTGGATATGTTTTGAGCAATAAAGAAATGATTGGAAAAAGTACACAATCTCCCATGTGATAArs11125229GGCAAGTCATCCTGCTCAGTGCCCTCAGAACATGCTTTTTTTCTTTCAGTATCTACAGTGCCTAGAACTGTGCCTGGACAAAGAAGACCTGAAGTACACA[A/T]TTGTTGAACTGAGTCTCTTTTAATGTCTAGTAAGCCTGGTGCTATAACTTTATCCTTATGCAGTCAGCAAATATTCGAATATACACTGAGAGAATCCCCArs9992168AGGTGGCTCCTATTTAACCAAGGGCAATTCTCTGGAGAAGGGGGCACCTGTTCATTATTATCCCCCAAACCTCACATCACCCAGAGGATGTGTACACTAA[C/T]TGGTACTAGGGAACTAGGCAGAGCACCAGTTGCATCCACTATAGTCCACTCTTCATCTACATGCTTCTCTCATTAAGTTCAGTCCATCCAGACACAGCTTrs7900002CCCTGCCTGAGGATTAATCCTTCCCTGCTACAGTCACACAACTGCCTCCTTCAGGGAGGGGAGAGTGCTCAGCTACGTGACCCAAAGTTCAGGATGGTAA[T/G]TGATGTCAAAAAGAGGAAGAAAGTTTGCATGTAGGTAACCAGGAGTGAGATCATGAGAAATGCAGGGTCTTACCCACATTTGCCCCATCTGTGTATTCAGrs11685586GCTGAGCTGCTGGCAGAGGGGAGGAGGCTGTGGGAACCAAGGAAGCTACCAAAGTGAACTTGGGTCTCCGAACTCACCACAGAAGCGGGGACTCCAGGAA[T/C]TCTGTGGCAGGTTGTTTCTTTCTCCCTCTACTTTATATGAAAAACACCTGCAGTGACCAACTCAAGACTATGAATGGTCATCACCGCACAATGACATGGTrs12158945GATGTATTCAAAATATATTTTTCTGCTTTACTATGTGATACTAATTGTAGCCCCTGTTCTGGGTTCCTTTTCTCATTTTCTTGCTTTCCTTTGCAGTAA[C/T]TGAGTTTTTAAAGTCATTCCACTTTTTCTTCTTTTAGTTCAAAAATATGCACTCTTTTATTCTAGTGGTTACCTTAGAAATTATAATATACATCATTGACrs12903747TACCAACAGCTGGGCAAAGTTCCAGGACAAAGTTTAGGGCAGGTTCCCAGGCACAGAAGCAAGTGGAGTTGAGGCCATTAAAAGCAGGGGTCTGGTATTG[A/C]ATTGGCCAAGCTGTATAATGTCCCGCAAGTTAGTGAACCTCTGCAAGCCTGAGTTTTCCACTATGTGAAATGAGTTCATCATAGTCCCTATCTCACAGGGrs11249671TGTGTGCCCAGAGCAGGGCTGGGCTCTCATAGCACAGCGGCGGCAGCACAGACCTTGCAGCCCTGTGGAGCTGTTATTCTAGTGTGGGAGGAAATGACCC[C/A]ATTGTCTCGACGGTGGTCCTATCAAAGAAGTCGCATAGGGTGACCTGGATGAGTACTGTTGGGAGCAGAGGCCAGGGAACTAGGCTCCACGCTGGGTAArs17079191AAGGAAGTGATGGGGAGGAAAATCATGCAAGGATAACTGTTATCTTGATTTCCCACCCTGAGATTGGGTGGAGGGGGAGCACAAACATACATTGGGGTAA[A/T]TAGAAATATATAGCAAATGCTACACTTAAGCTTGGAGACCATGGTCTTTCCAATTCAGTGAATTTTTTTTTTTTGAAATGGCATTCAAACTTTGTTTTGCrs10832561AAATAAGCACAGACTGGATTTTAATTTTCTAAACTGATGTGCCTTTTTAAATTGAATACAGAATAGTCTTCAAATGGAAAGGGCCACTTTTTTTTACTGA[A/T]TTAATGTGAAACATACTACCACTTTATTGCTAGATTAAAATGTTAGACTAGAAGAAATAACCTAGTAGTTTGTCTCATAATATCAATTGAATTATATGAArs11563997CATTAACCGGATATAAACTTCTTATTGGCCTTCTTGGGACTCAAATGCTGTACTATTCATCGAGTAAGAGCTTAGTAAACTTGAAGAGAATAAATGAATA[C/A]ATTGATATAAAAGCCTTTTATGTTTAAGTGTTTTTAAATCTAATAGTGATTCTAAAAAAGAGAGGGGTAAATGATGTGTATTTTGCTCTAAGATTTCCAArs10754776AGAAAATGAACCTTAATCTAAACATCACAACTCATACAAAAAGTAACTCAAAATAGATGATGGACTGTAAAATGTAAAACTCTAAGACTTTTAGAAAAAA[T/A]TCTATATGAGAAAGTATTCAGGATGTAAGGCTAGGCAAGGCATTCTTAGACTTGATATCAAAGAGCATGACCCCAAAAAAGAAAAAAATTGATAAATTATrs7985274GCTCTTCCCTAAGGCCTCATCAGAACGAGGCCTTTATACCACAGAGGACACACACACCACACAGACTGGACATCTCAGAGGGCCCATGGCATGTTTTCAA[G/A]TTGCGGAGAGCAAAAGAGAGGCCATAGTTAGGACTGATCATAGTTCATCCTCAATCGTGTCAATGAGTGCAGGTAAGCCAGGCTGTAGAAAAACCAAAGArs10234234TTGTAGAGAAATAAAACAGTGGCTGAAGGGGGTGTGCATGTCGAGAGAGAGAGAGTTTGAGAAGGGAAGTGTTGTAGCATGGTTGTATGTGGATGGGATT[A/G]ATTCAGTCAAGAGGGAAAAACTGATGATGCAGGGAAAAGAAGGATAGTTATGAAAGTGTTATCCTTCATAAGTGAGAGGGAATGGGATCTTGTGCACAAGrs9314663ACCTCCACCTGGTGGGGCCTGTCAGTGTACCACAGGTCTACCTTGATTTCAAGTCCATCTCCTAATAATGAAGTAAAATGTTTTTCCCTGCATTGAAGAA[A/C]TTTCCAGTGTCTTGGATGGGGGAGCTTAAGGAGCAGATGCTCATTCTTGGGGTATGGAGGTGATAACTTGTAGGCAGACTGTTCCTAGGAACACACGTAArs10021843AAACAGTGCAGTGCAGTTGTTGATCTCAGCTGTTTTAATGCATGAAACATGTTAAAACATGTCAGTATTAACTGTGAACTTTTTTTGCAAGGGAGGAAAA[C/T]TGAGATAATATTCCTTTGAATCATGAACAACAAGTGGTTGATAAGTGCTATATCCCTGGCCAGCTTTTTTGTGTTGCTTCATAGCTGAGCCACATCAGTTrs11773909TTTAGAAACTGAAACTAAGTATATCTGATGTTGCTTTTAGGAAACAAGTAAATGAGGTCCTAAAAAGTTAAACTGTGACCATATTTTCTTTCCTTTTTCT[A/C]ATTTCTCCTTGGGCCATTTCCAAAAAGCCCTAATACCCCGACTGATAGAAATGGATACCTTGCTGTGCACTGGTACTACTGTGATTCATGGAAAGCTGATrs11227624GCAGATAACCACAGTGGGAGGGAGGCTTCCCCTGATGGGCCAGCAGGGTTAGGGCACTCTCATTACCCGCTGCCTGTGCAGCAATGATCACAGCTATAAT[T/C]GAACAGGGAATGGCCTTCTGCCAGTCCCCCCTGATGACAGGAGATGGCTGAGGCCTTCTCCCTGCTGTGTCTCCAGCATGAAGCATGCGGCCTAGAACACrs9838013TGTGAGAGCAAGGATTCTTTATCTACATATAAAATAAAACAAAAGTGGAACCATATTTTTGTCCCCAAACATCCCTTTGATACTACCATTGAGGTTTCAC[A/C]ATTAGGACAGTTTTCTTCCAGCACCCTCACTAAACGACACCCCTCTACTCTCATCTTGCACAATTCCCTTCCTTCCTCTCCAGCAAACATTCCTCTATTTrs9929404CAACCATGTTTACCAAATGATACTAAACAATTGATAAGATCATCTCCACATGGATAACAGCTGCTTATGGAGATGAGTAAGAGCAGGTGAAATGTTTCTA[T/A]TTCTATTCATACATGAGCAGATTAATAGAGAGCTAAAATGGTGTTCAGGGTCTTATGAGTAGCACTTTTGGTTAGGGTTTTCCTGTTAACATCCATTATArs13255815GGGTTTCATGTACGTGTGGATGGAGGTTGGCTCAGAGATGTTTCCACATTTCCAGCTCTGACAGCTGTTGGTAATAGCTACAGCCCTGGTCCCCTGGAAT[T/C]CGCTTCCCTGCCTGGCCTGACCCTGCGCTGACAGTCAGCTCTTCTCAAACAAGCAGTCTCAATGATGATAAGCATCTCCTTGGAAGGAGAAGCTTCGAAGrs9987005GGTAAAAAATTAAGCTTGCATTTCCTTTTTACACAGAAGCTCTTCCACTAATTCAAGCCAATACATTTACAATAGAACATGCCAGAAAGTGCCACAAAAT[T/A]TCAATAACAGGCAACACCACTAGGCTTCAGTGACCACTGATTTCATCCTCCTTCTCCTATATTCTTTCCTATAGTCCTTATACATCAATGTCATGGACTArs11635372CCACAGGAAACTTCACAAAGGTTTACGTACAGAAGCATTTGGGGCCATGTCTGTCTTGGCTATGGGGACAGGTGGGGCTAAGCCGGCATCTCTGCTGTCA[G/A]TTGCCAGACTGCAGAGAGAGGCCCTTGCCTCCTTCCACAAGGTGTTTCCAATAAAGGGGACATATTTCCTTCGTTAGAAATAAACACAGACTGACAATATrs12674093GAGGAAATGGCCATTTCTGAGGTGCTCAGAACCACAGGCTCACCCCTTTCACAGGGTTAGGATGGGAGCTGTTACAGGGAGTTTCCTGTACTTTAAAAAA[G/T]TTAAACAACAGAATCCAGCCTTTGCTAGCTTTGGGTACTGTAAATGATTTACTGTAACATAAAACACATCGAGTGAGAAAAATATAGAATAAGTTTTTTCrs10260483TTTAAGTGTCGTCCAAAAGAGATTAGTATTGGTCATAACATGGACTCTAAAGCCACCATTTAAATGAAGCATGTAAAAAAGAATATTCTAGTACACAAAA[G/A]TTATTAATGGCCTAGAATGACCTCCTTCTCACTCATATGATGCAAAGAATAAAGTATATAAAAATGTTTGTTACAATGGCTATCCATAAAAAAGAAAACCrs11759755CATTTAAAAATGTGTTTATCAAAAGACACTGTTAAGATTATGAAAAGGCAATCCACACGGTGAAAGAAGATATTCAAAATACATATATTCAACAAAGAAT[T/G]TATATCCAGTATATAAACACACACACACACACACACACCCTACAGATTAATAAGAACAAAGACAATCCAACAGCAAAAAACAATAGGAAATTATGAAAGTrs12783667GTCCCTGAAGATGTGTTGTTGAGAATGGATGACAAACAGTTCAGGTCAACCTTGAGTAAGTGTGAGGAAAAAATAAAAATAAATAAATGAAGGATGTAAT[T/C]GGGCTCCTCTCCTGGAGACTGAAAAGTAAGGACTGGCATGGAAATCTTTGATTTTTGGCAGTATATCATTATCTTTAGAGGTCTAGAAAAAGTGCCTACGrs9692857TTTACAACAGCATCCAAAAGGATAAGCTACTTCGGAATAAATTTAACCAATGAGGTAGGAAACGTGTACACTGAAAACTATAAAGCATTGCTAAAAGAAT[C/T]TAAAGATGATACAAATGAAAGAAAAGACATCCTGTTTTCATGGATTGGAAGACTTAATATTGTTAAGGTGTCAATACTATTGATACAGTTTGGATCTATGrs9428474CCTCCTGCCCGGTCCATAGAAAAATTGTCTTCCATGAAATCGATCCCTGGTGCCCAAAAGTTTGGAGGCCACTGGATTAAAGGAGACAATGTATGTAAAT[T/C]TTGGCTTATAATAAGTTCTTGGAAAGTGCTAGCTGTGTCTTATCACTGATTATAGTATCCCAATCAAACCTTGACACTTGGGTTAGGATTATTTATTTCCrs16830436TGTGTGTGTGTGTGTGTGTGCGCGCGCGCGTGTGTGTGTGTGTGTCCACTGGCCTTTTCAAAGTCTCTCTTTTGTTTTGCAACTTTGGCTTTATTTATAA[T/G]TTAAATCTAGACATTTCTTCTTGTGAAGTCCATGCACGCCACTCTTGGGACATCCCTATGTTGCAGCAGAGGATAAAAATGGAAAATTCAGGGTCCTTAArs8063107CTGAATTCCTGTAAAGAAAGGAGACTCATATCCTGAAGAATGAAGACATCAAAAGCAAGGTGCTGTGGCAAGTTAGCCCTTGGTGGAGGGTTTTTCACAA[C/T]TGGATATCCTGCTGTGTAGAACTGAATACCCACAGCAGGGTTATTCAGGCAGCTCCAGGGATGAGAGAAAGTGTCTTGACTGATACATAATTTATCTGTCrs9818611AGAGTTACTGTTGCTCCACGTCCTACCAGCATTTGGTGTCAGTGTTCTGGATGTTGGCCATTCTAATAAGTATGTAGTGCTATCTCATTGTTGTTTGAAA[C/T]TGTATTTCCCAGATGCATATGATGTGGAACGTCTTCTCATATGCTAACATGCCATCTGTATATCTTCCTTGGGGTGTCTGCTAAGGTCTTTTGCCCAATGrs10840805TACTTGTTAATACTCCAATTACTTCCCAGATTAAGAGATTTGTTTCTCTACAACAAATATTTGTACCTACCTTGCTCTGAGAAACAGCCTGCACTGTGAA[C/T]TCATTTTATCAACAACAAGACTGCTTAAAAGCAGGAAGAAAAAGCCATAAAAAATGATGAGTTCACGTCCTTTGTAGGGACATGGATGATACTGGAAATCrs10421748GAACCCCTTCCTTGCCCCTAGACAAGCCACAGCTGACCTGCTGAGCAGCCTGGAGGACCTGGAGCTCAGCAACCGACGTCTGGTTGGGGAGAATGCCAAA[T/C]TGCAGCGGAGCATGGAGACAGCTGAGGAGGGGTCAGCACGCCTTGGGGAGGAGATCTTGGCTCTGCGTAAGCAGCTTCACAGGTGGGCTGGATGCCACACrs10139699ACTAAACAAATGTATTAAATGTTCCTGGCTCTGTACACCATCCTTTAGGTAGAGAATAATGGCAGGCATTTGGGTGTTTCTCAGGAGTTCCCAGCAGAAT[C/T]GACTACCTTTGCCCAGAGCAGTAATCTTAGTAATGCACACACAAGTTGTCTTTTTCTCCTCTCCTGCATCGTTAAATAAACTACAAATATATGAGTAGAArs12107918ATGAAATGGATTCACATTTTTAATGTTCTATGTAATTACTTATCATTGTTGTTTTAATAGGGAAAGTATTGGTTATATAAATAGCCAAGAAAACAGCCAA[C/T]TGAGACTTTTCTTCCTAGATTACCTTGGTTATATCAGTGCTTCTGGGTGTGGTCACTGATATTCTACAGCAGAAACAGCTAGTGGGGTCCCCAACTAAAGrs10884498GATGGCATATGGAGAGGACTTACAAAAGGGCTTCGGAAATATTTATTATTATTATACAATAATACATGATATTTTGTGACGGTTAATACTGAGTGTCAAA[T/A]TGATTTGATTGTAGAATGCCAAGTATTGATCCTGGGTGTGTCTGTAAGGGTGTTGTCAAAGGTGATTAACATTTGAGTCAGTGGGCTGGGAAAGGCAGACrs10822434AGAATGTATTTATTGATCTGTGATATCTATCCATACACCAATAGTAACTATTTTATATAAACTACTTTTTTGAAAAGTCTTGACATAAGGTAGTATAAAT[T/C]CTGTTGCTCTTCTCTGTTTCAGTATTTCCTTTGCAACCCTCTTTAAGATTGCCTTTCACTTCTATGTAAGTTCTCAAAAGAGGTTGTTAATTTTAATAAArs12607335AATATAAGTGGAATCATAAAATAGGTGGTCTTTTTTGGCTGGATTCTTTAATTTATCAAAATGCTTAGAAGGTTCATTTATGTGGTAGCATGTAGCAGTA[G/A]TTATTTCCTTTTGTTGTCAGATAATATCCATTGCCTCAATAGACCACATTTTCTTCTCAATTTATCACTTGATAGACATTTGAATTATTTATACTTTTTGrs7915178CAGAGCTATCACCTAAAAGCATCACATGGACATTTAAAATTCTCAGTAGAGCATTTTTTCCTTCTAATGAAGCTTTCCTAAACCTGTGACATTGGTTTAA[T/C]TTGTGCAGGAGTTTCCTCCTTGTATTTGTTTAAATGCCCCCAGAAGCTCGGAAAGCAGGAAGTGGTTTGAAGGGGATTCAGACAAGGTTAGCTGGGGAGGrs10953770GTACAGTGAAAGCACTTCAAAATCTTTCAGGTGTAATCATAAGAAATTATTTATCTTAGGATTCTTGATATATTACATCGAAATCAAGGTTTATGTTATA[T/A]TTGAGTAAAGTTTTCAAGGATGAAAACGATTTTGCCTATTTTTTTCTGAAGAATTACAAACACCTGCTTCTTTCATCTTCCTTTGACACTCTGTTCCTGArs11099210GCTCTGGACCCAGCCACGCTGGGAGGGAAACCACCTGATTTCAGGTACAGAACCACTCTCATGTACCCTCTCTGCTGAGAGTTATTCCATCACTCAATAA[A/C]ATTCTTCTCTGCCCTCCTCACCCCTTGATTGTCAGTGTAACCTCACTCTTCTTGGACGCTGAACAAGAACTGAGGAACTGCTGAATGCAGGTACAGCTGTrs10785736TACTTCAAATAACATCTACACTTTTTAAAGAAGAAGATTCAATCTCAGAGAAACTGGTTTGGTTTCTCAGCTGGGAATATTTATTTGGTCATACTAAACA[A/G]TTGAGCCAGTGGATCAGCAGTAGCTGATTGCAAGATTCTTAAGTAGACACACATTACATTTCGTAGGGGATCAAAATATGTCATTCTCAAGTATGCTAATrs11221881CCATCTCTAATTTCCGGGAGATTTATAATTTGTTTGTATTATTTTGTGAATCATCCGTTCATGTCTTCTGCCTATTCTTCTACGGTCTTTTTCTTATCAA[T/C]TTGTAAAGACTCTAATGTAATAGCCAACTGCTACAAGCATGTTTCTGATTTGTTGTTTACCTTTTGATGTTCTTGATATTAAAAGATGCTTATATAGCTGrs11727770CTCCACATCTGTCTACTTGCTTGTTGACTATCTTACCCCCTTAGGCTATAAGTACTCACTGATCTGTCTCAAGTGTCTGGTTCATAGTTAAAAGTCAATA[A/C]TTACGTGATGAATGAATGAATAGATGGAAAAATCAATGGATGGGTGGATGGATGATCTTTACAGATTAACTTGAACCAGATCATGTAAGGAGCTGTTTAArs10102733TTTAGCTTCATGATTTAACAGGAATAGTGTGAGGTAAAATGACATGAGTCACTTAAAGCCTTTCAGAAGGAGAAGTACCAGCCTTGATGTGGGGAAAAAA[T/C]TGGTCATGGTGGCTCACACGTGTAATCCTAGCACTTTGGGAGGCCGAGATGGGCGAATCACAAGGTCAGGAGTTCGAAACCAGTCTGGCCAACATGATGArs10030074CAGGACTTTGGGAGGCCAAGGCAGGTGGATCACCTGAGGTCAGGAGTTCGAGACCGGCCTGACCAATATGGAGAAACCTGTCTCTACTAAAAATAAGAAA[G/A]TTAGCCTGGCCTGGTGGTGTGGGACTGTAGTCCCAGATACTCGGGAGGCTGAGACAGAAAAACTACTTGAACCCGGGAGGTGGAGGTTGCAACGAGCGGArs10510379TTAGCCAGGATGGTCTTGACCTCCTGAAATTGTCATTATTTGCTTTTAATGTGGATTGCTTTTATGAGAATAACTATGAGCTCATGGATTTTATATAGTA[G/A]TTGTCACGCATGTCCGTGTGAAGAGAGTCCACCAACAGGCTTTGTGTGAGCAACAAGGTTGTTTATTTCACCTGGGTGCAGGCAGGCTGAGTCCAAAAAArs13110085TGCTGTGGTTAGGAGGTATAACTTGGTTAAGTGTTTCTACCCACGCGTAGGCTATGGTTTACATAGCCTATGCACACATAGCCTATGTGTGCATAGTTTA[C/A]ATTTCCTACCAGCCCCCCAAAAAGGGAACACTTGCTTTTCTTATCAACTTGCCCAAGATGTGGGGTAGAAGGGAAGGGGCAAGTGGTAGAGCTCGCAAGCrs13269702TACTGTTCTCAAAAGGCAAAGTCCTGTGTAGTTCATGACTTCTGTGGACCATACAGAGATAAAAATAAGAATGGAAAGTGATGAGATTTCTATCATACAA[A/T]TGTCATTTCCTGTGAAAAGGCAAAGATGATTTCAAATAGTGAACAAGCCTAGAAAGTTTTTAAGGGGCTTTGGAACATGATAGAGACACACAATCAGACArs9312864CTGGCCTACAATTTTTTTAAAGTGATAACATGAAGAATAAAAAAGCTGACAAACTGTTCCAGATTAAAGGTAAGTAAAAAATCATTATCACTAAAGGCAA[T/C]TTGAGCTTTCGATTTGGCTCAGGATTGTGGGGAAAGGAGGTGGGGTGGGGAAAAGAGGTGGAGGACATGAGTTGCCACAAAATTATTGAAACAATTGGTGrs9787011GATAGTCCTTAGGCCTTGAAGTCATTAGAGGTGCAATGCTGTAGGGCCAGAATGGGTAAAGGAGGGAAGGTGGGATAAACAAGGGGGTTTGTGGTGAAGA[C/A]ATTCACTTGAAGGGGCACTTAACATGTTATCTGACCTGTGATAAGTGCTCAGTATTTGCCAATGGATGAATTATGACTGAATGAATAAAGTCACAACCTGrs9555581GAATATCTTTACATTGATTTCCTGATGCGCTACATATCACTGCATGCATTGAAACCTGGGATACACAAAAAAGTTATGCTGAGGTATCATCTCTAAGAAT[C/T]CAATACAGGAGTGAGGTCGAGATTGCCTTTTGAGAGTAAATCCAGAAGCCAACTTAATAGATCAGAGCAGAATTAAGGCAAAATTACTTTAGGATAATGGrs8016543AATGAGGATGATGAAGGGAAGTGTCTGTGCAGGGCTTTTAACCCTTCAGGAGTTTGGCCCAGTTCATCAGAGAGAAAAGGGAGTAGGTACTTGTCACAAA[G/A]TTTGGCTTCAGTATCAGGTTTTCATATGGCTGGCTGGAACCTGTATAAGGACCCAGGAATGGATAGAGCTACTTTGTAATGAGAGACTTTGACACATTTGrs13155942CTCTGTGATTTTAGATTGTGGGTTTATATTTGCTGGCATGCTATCTCTGCGGTTTCTTTTGAGGACTGGGTTGAGGGTACCTTTCTTTCTCCAGGGATGA[T/A]TTGTGTTTGCTTTTGTCAGACACTGAGCACTACTGACATGAATCCTCTTAAAAATACAATGGTCAGCCATATAAACTACGTAAACAGTGTAGATTCAGTTrs11655850TGCACAGTCACCCTGGAGAGAACCATGCCACAAGCCCAGATCCAGGACACACTGTCATGCAGGCTTCCTTCCCCACCACTCAGGAAAGCAAACTGCTACA[A/C]TTAAAAAGGAACAAGGGCAAGTCTGGTGCTGCTCTTCACTGGGATTTTTTTCTTTTTTTTTTTTTTTTAAGACAGAGTCTCACTCTGCCATCAGGCTGCArs13331222ATAAATATATAATTTAACAAAAGAGAATAACAAGAACGAAGTAAAGGAATACATGTGGGTATGTGTGTATCTATGTAAAAATGGAGAGCCATGAGTGAAA[T/C]TGTATACCAAAGGAAGCAACGTATATTCTTAAAAAGGAAAAAAAAAAGACATGAGAATGCATTGGTCTTCCGTGAAATGTAGCTACTGTAAGGTTTTTATrs10110766GGGCATCTTTCAGTTAGTACGTGGTACTGGACAAGAGATCAGGTTGAACTGTAAGGCTCTAGTTTTCAGCAGACTCATGGTCCTGGGAGAAAGAAAACAA[C/T]TGGACAGGGGGCTATCAAGGTAGCCAGGTTTTGAGGGGACTCAGTTCAGGAGAAAAGAACTGGAAAGCAGGTCCTGTGCTGCTTTTCTCCTTGAGAAATTrs9554894CATTCCTTTCCATTACATACTTTCTTTGTCGACCTGAGTTTTCAGCCGTTGCTGAAATAAAAGCAAGTATTGCACAAGAATCAGTTTGGTGTTCCATCCA[A/C]TTCCAAAGTTTGAGTTGTGTCATGCCCAACAGGCAAACACACCTCACTCAGTAATTGTGGTTAAGAATGAAATAGGCGCAGTGGCTCACGCCTGTAATCCrs17152417AAACAAACAAACAAACAAAAAAACCCATAATTCAGCCCACCAGTGGCCTCAGGTTACTGTGTGTACAAGGTGTTTGTGGGATATTTCTGGTCTCCCACAA[T/C]TTCAGCTGATGTCCAGAGTTAAAGGGCTCTAAGTAAGTACCCCACCTTCTATAAAGTGTTGCTAAGGAAAGCCCTCAATGCTAAGGCTTTGATACAAAATrs17156383ATAGACATTCACTAAGATATTGCATCTATGAAAAATAATTACACGCTATGTAAAAGTAGCAATAAAAATAAAAAAAGCTACTGAAAATGAAAATGTATAA[T/C]TGACAAACATAAACTGCATATGAACTTTGGAAGAGTAAATAAAGTATCCTGGAATATAGAACAAAATAATATAGAGTAAAAAAAAAGGAAAAATCTTAGArs11017936GGAACCAAGTCCCATCATTGCAATATCTCTCTGGATTCCATTGTAATCCATTTCAGACGCAGCCACACGTGTTCATGAACTCATCAATGCAATCTGGAAA[T/C]TTGACTTTGGCTTGTGATCTCTGACATTTTGATGTTTTAAAGTGGGTTTTCTGGAGTGGAGTCTTGGGCCTCCCTCTCACACTTACGGAGTCTTCCTATGrs10777944CTATTTGCTTGTGTCCCTTCATCTCCTGCTGGACCATGAGCTCCTGGAGAGCAGGGATGTGTGTCTAATGCATGGCAGGCACTCTATCAATACAGGAATG[C/A]ATTTTATGTGGAATCTGACTTTTTTCCTCAGATGTGGAAGCACGCAGCAACAAACATATGTCGTGAATCAAAAACCGGGACATAAAGCCTCACACAGGGTrs10278812TTATTTGTGAGCAGTATTTTAGTTTTAAATGGTAGATATTAAGCCTGTACAATGATATTCAAACAATGGTATATTGAATGGATAGAAGAATCTGTCATAA[A/T]ATTAGAGTAATGGTTTGAAAAACCAATGTTTGTGGAGATAGCAGTCAGGGTAGTTATGGGGAGAACAGAGACTAGAAGCTGAAATTACAAAGTGATCAAGrs10784847GGCAAGACAAAGGAAAGGGAGTTCAGCCTTGTAGGGGTGGTAAATTGTGGATTTTCCTGGTATGAAAGAGTGAAGGGAGGACGTTTTCTTAAACAAAAAT[T/G]TATGCCCTGCTTTCAAGCAAGTAGGGGGAGGGCACAGAGCTTTTCTGTGCCTGCTATTTCTTGATTGCCTTCAGCTTAAAATAATTCTTATGTCAAAGAGrs10179379TACACAGGGCAGACATGATGGTGGCTTGGCCCAGTGTGGTGGCAGCAAGGGTAGTAGGAAGTGGTTAGATTCTGGTTATATGTTAAAGATAGAGCACCAG[C/A]ATTTCCGGACAGATTGGATGGGAGGTGTCACTAAAACAGAAATCCAGGGATAACTCTGAGGTGTTTGGCCTGAGTTATTAGAATGATAATATTATATTTArs17074340AATGTGGTGGCCTCACACTAAGACGTAGAGAAGAAGAGAACCTAGAGTCAATGAAGCTCATAAAATGCTACTCCAACAGAGGAGGTGACATAAGTAAGTA[A/T]TTCCATGGGAGAGGGAGGTCAGCAGTGGGGATAATGAAGAAAGGAATATTATAAATACATTTTGATGGAAAAATGTAAAAGGATAAGTCATTAATTCCCTrs1297215ATTTGCTATCTCTGGATATCTAGCTTATTTCTAAAAACCTCTAGTGACCATGAACTATCTTCCAAGGTGGTCTTTTGGAGACGGATGGCTCTGGGTTCAA[T/C]TTATTCCGGCTCTACCATTTACCAACTCTTTGATCATAGGAAAGTTGGCTACTCTTGAAAGTTTATCATTATTAAACGTGCAAAAGCACTAATACCTGTTrs1041409ACCATATTAGTAAGTCTCCCCTGCATTATGGTGTACTGTTAGTGTGTCACTCATATCATATCAGATTCCTTAAACATTTGTTTGCATAAAGTCCCCATGT[A/G]ATTCTATTCCCCATAGTAAGTACCTGCTTCTCTAGCACCATGTACTATGTACTATGCACAAGTAGCCAGAATCAGATTTGTCTACAGAATTGGAGAACTArs2826737TTTACCAGGAATTGTGATACTTCATTTATACACATACTTTATTTAATCCTTACCATGACCATAGATGACTTACATATGCTAAGAGCCAGGACTCTAGTCC[G/A]ATTCAAATCTGTCTGACCCCAGAATCCTTAGCATTTTCAATGTGTTTCTGGAAATAGCCTTACCATAAACCGCAGTTGCACTTTTTACCACCTAATGTGTrs2834712GGACTTACAGTCTCATTCAGGAAGACCTTGACAAACAAATGCTAACATAAAAACCACCAGACTGCTATTTAGCCATTCTGTCTGGGATGACTATATTAAT[T/C]ATTTTATGACAGCGTTTCTTTCCTTCTGAATGGTTGTTACCAGCGAGGTACCTTTTGCTCAATGTTTGCTTAAAGACATGTCTATATATTATCTGGCAAG

Conditions Used for Testing

PCR

PCR was performed with or without the addition of Tsp509I to the PCRcocktail mix as indicated in Table 12. PCR cycling was performed for allsamples with the cycling conditions in Table 13 to allow Tsp509Idigestion of the DNA immediately prior to PCR amplification and in asingle tube. This was used even if there was no Tsp509I added to thecocktail.

TABLE 12 Volume per Reagents Final Conc reaction (uL) Water n/a 3.12510x PCR Buffer 1.25x 3.125 MgCl₂ (25 mM*) 1.625 mM 1.625 PCR NucleotideMix (ACGU) 0.2 mM 0.5 F/R Primer mix (0.5uM) 0.1 μM 5 10U/ul Tsp509I 0or 0.02U/ul 0 or 0.05 1U/μl Uracil-DNA-Glycosylase 0.05U/ul 0.625HotStar Taq (5U/uL) 0.2U/ul 1 Total volume n/a 15 DNA - added separatelyvaries 10

TABLE 13 30 C. 10 min UNG digestion temperature 65 C. 15 Tsp509Idigestion temperature 94 C. 15 min Taq activation 94 C. 20 sec 58 C. 30sec {close oversize bracket} 45 cycles - Amplification 72 C.  1 min 72C.  3 min Final extension  4 C. Forever Storage

SAP

SAP dephosphorylation was carried out with standard conditions includingthe SAP cocktail preparation below in Table 14.

TABLE 14 Table 14. SAP Cocktail preparation SAP Mix Reagent Volume perReaction Nanopure Water 2.95 μl SAP Buffer 0.34 μl Shrimp AlkalinePhosphatase (SAP) 0.71 μl (1.7U/uL) Total Volume   4 μl

TypePLEX Extend

TypePLEX Extend reaction was carried out with standard conditionsincluding extend cocktail preparation below in Table 15.

TABLE 15 Table 15. Extend Cocktail Preparation Volume per Extend ReagentReaction Water (HPLC grade) 1.24 μl TypePLEX buffer (10x)  0.4 μlTypePLEX Termination Mix  0.4 μl Extend Primer Mix (3-tiered 5-15uMstock conc) 1.88 μl Thermosequenase (32U/uL) 0.08 μl Total Volume   4 μl

Digestion of Heterozygous SNPs in Genomic DNA

CEPH genomic DNA obtained from the Coriell collection was used to testthe ability of Tsp509I to specifically digest one allele of each SNP.The informative allele peak area ratios of DNAs heterozygous for theindicated SNPs were determined. The informative allele, alternativelycalled the target allele, is defined as the allele NOT recognized byTsp509I enzyme. Tsp509I treatment significantly increased the peak arearatio. With no Tsp509I treatment, heterozygous DNAs show median alleleratios ranging from 0.4-0.6 depending on the SNP. After Tsp509Itreatment, for the majority of heterozygous DNAs, the median peak arearatio is above 0.8 with many peak area ratios at 1.0. Peak area ratiosof 1.0 indicate that there is no detectable non-informative (i.e.,non-target) allele peak area present.

2% Mixture Model

A DNA mixture model was prepared from CEPH genomic DNA obtained from theCoriell collection. The DNA mixture model was used to test the abilityof Tsp509I to enhance the detection of one allele of a SNP when presentat a low fractional concentration. Briefly, the DNA mixture modelcomprises 47 unique child/maternal DNA pairs mixed together such thatthe child's DNA (the low fractional concentration DNA) is present atonly 2% of the total DNA. For the studies here, DNA was added to the PCRsuch that there were 20 genomic copies of the low fractionalconcentration DNA and 980 copies of the high fractional concentrationDNA in each PCR. In these mixture studies, not all DNA pairs will yieldinformative data for every SNP. Informative data can only be obtainedfor a SNP when the maternal genotype is homozygous for thenon-informative allele and the child's genotype is heterozygous for theSNP. With no Tsp509I treatment, potentially informative DNA mixturesshow median informative peak area ratios at background levels. AfterTsp509I treatment, the majority of DNA mixtures with potentiallyinformative genotype combinations for the indicated SNP show median peakarea ratios above 0.5 with many peak area ratios at 1.0. Peak arearatios of 1.0 indicate that there is no detectable non-informativeallele peak area present. This indicates the utility of the multiplexedSNPs to detect a low fractional concentration DNA present at least aslow as 2% of the total DNA present and at levels as low as 20 genomiccopies of DNA.

Detection of Low Fractional Concentration DNA

Modified versions of multiplexes 2, 5, and 6 with a total of 95 SNPassays (see Table 16) were tested for their ability to detect a lowfractional concentration DNA. Sample test groups included:

-   -   1) Maternal only genomic DNA used in DNA mixture models    -   2) 2% DNA mixtures of child/maternal genomic DNA    -   3) Maternal PBMC DNA (pairing to the plasma DNA below)    -   4) Maternal plasma DNA previously shown positive for 8        Y-chromosomal markers indicating the presence of male fetal DNA        (pairing to PBMC DNA above)

For the comparison, each of the above sample types was digested withTsp509I prior to genotyping with the TypePLEX extend assay. Separately,maternal genotypes from undigested maternal DNA was determined toidentify potentially informative SNPs for each sample. For thisanalysis, no genotype information obtained directly from child genomicDNA or fetal genomic DNA was used.

With 95 SNP genotype assays, one would expect to have 3 or moreinformative genotype combinations in ˜99.9% of cases with biologicallyrelated maternal and child genotypes.

Therefore, detection of at least 3 informative SNP alleles present in aTsp509I digested sample that are not present in an undigested maternalonly DNA sample should allow detection of a low fractional concentrationDNA. Increasing this required number of detected informative SNP allelesto greater than 3 will likely increase the specificity but at theexpense of sensitivity.

In prior studies with the DNA mixtures, it was noted that in the Tsp509Idigested samples, background levels of informative allele peak areacould lead to artificially high detection of an informative allele peakarea ratio. Therefore, preliminary threshold criteria were establishedto improve the accuracy of detecting informative SNP alleles arisingfrom low fractional concentration DNA. In the data here, thesethresholds are defined as follows:

-   -   1) Informative allele peak area ratios must be at least 0.4        greater in the digested DNA mixture or digested maternal plasma        DNA sample versus the matching undigested maternal sample.        -   and    -   2) There must be greater than 15% primer extension product        generated for the SNP.

The criteria used here to determine the presence or absence of aninformative SNP allele are preliminary and are only exemplary.Additionally, individual SNP assays within the multiplexes may havetheir own criteria. Alteration of these criteria can have significantimpact on the detection of informative SNP alleles in either a positiveor negative manner.

As can be seen in FIG. 10, there is a clear delineation between mixedDNAs and maternal only DNA in the DNA mixture model, where at most 3informative SNP alleles (as defined by the criteria above) are detectedin maternal only DNA and 6-18 informative SNP alleles are detected ineach of the DNA mixtures containing 20/980 genomic copies ofchild/maternal DNA. In the plasma sample testing, this delineation,while not as clear as in the DNA mixture model, is still present betweenmaternal PBMC DNA and maternal plasma DNA. Here, maternal PBMC DNA showsat most 3 informative SNP alleles detected while the maternal plasmaDNAs show 4-19 informative SNP alleles detected. The dashed linesrepresents a possible cut-off value for informative and non-informativealleles. These data provide an evaluation of the utility of the methodto detect low fractional concentration DNA.

Detection of Fetal Identifier Alleles in Maternal Plasma

The ability to detect fetal identifier alleles in maternal plasma DNAand non-pregnant female plasma DNA was compared. Ninety-two of the fetalidentifier SNPs in Table 16 in 3 multiplexes were assayed by genotypingbuffy coat, PBMC or whole blood genomic DNA from plasma samples. Thesamples were analyzed in parallel with and without Tsp509I digestion,and they were subsequently genotyped for the same SNPs. Genotypemeasurement was performed on the MassARRAY® system. A fetal identifierallele was counted as ‘detected’ if the undigested genomic DNA washomozygous for the cleavable SNP allele and the matching plasma DNAsample showed the presence of the non-cleavable SNP allele afterdigestion of the plasma DNA with Tsp509I.

FIG. 11 shows the results from the 117 plasma samples tested for the 92SNPs. The x-axis of the dot plot above indicates the number of fetalidentifier alleles detected in a plasma DNA sample. Each dot in the dotplot field represents a sample. The top portion of the panel comprises27 non-pregnant plasma samples. The bottom portion of the panelcomprises 90 pregnant, maternal plasma samples. The legend providessample type and fetal sex (if known).

As expected, the fetal identifier alleles were detected in the pregnantmaternal samples and not the non-pregnant plasma samples. As the numberof SNPs tested increases, the probability of the number of informativeSNPs also increases. This is shown graphically in FIG. 12. The FIG. 12graph shows the probability of the number of informative SNPs for eachof the selected thresholds (1-6, shown each with a different color) atincreasing numbers of total SNPs assayed. For example, if 90 SNPs areassayed, the probability of at least 4 SNPs being informative is almost100%.

TABLE 16 MP2.1 MP5.1 MP6.1 rs11835780 rs748773 rs7323716 rs4311632rs6488494 rs9652080 rs13110085 rs1363267 rs10785736 rs13269702rs10840805 rs7356482 rs1797700 rs2723307 rs7831906 rs2993531 rs4764597rs9818611 rs1885121 rs4589569 rs12903747 rs1372688 rs2820107 rs7320201rs1904161 rs6766358 rs4869315 rs1720839 rs12675087 rs3913810 rs10901705rs7689368 rs6542638 rs6582294 rs725849 rs12450474 rs7144509 rs7900002rs1346718 rs10234234 rs910500 rs1503660 rs8016543 rs4489023 rs2007475rs11221881 rs1916803 rs683262 rs13155942 rs10260483 rs10110766 rs494220rs4488809 rs1041409 rs3912319 rs4533845 rs11099210 rs9428474 rs3816551rs10754776 rs9929404 rs6556642 rs4130306 rs7294836 rs7205009 rs2734574rs4673821 rs12674093 rs1401454 rs614004 rs2322301 rs9285190 rs1444647rs7741525 rs179596 rs7818415 rs17074340 rs331893 rs12007 rs2462049rs9787011 rs9356029 rs10806232 rs6569474 rs11105611 rs9989393 rs10898954rs12107918 rs6043856 rs664358 rs263025 rs1593443 rs6142841 rs1342995rs273172

The entirety of each patent, patent application, publication anddocument referenced herein hereby is incorporated by reference. Citationof the above patents, patent applications, publications and documents isnot an admission that any of the foregoing is pertinent prior art, nordoes it constitute any admission as to the contents or date of thesepublications or documents.

Modifications may be made to the foregoing without departing from thebasic aspects of the invention. Although the invention has beendescribed in substantial detail with reference to one or more specificembodiments, those of ordinary skill in the art will recognize thatchanges may be made to the embodiments specifically disclosed in thisapplication, yet these modifications and improvements are within thescope and spirit of the invention.

The invention illustratively described herein suitably may be practicedin the absence of any element(s) not specifically disclosed herein.Thus, for example, in each instance herein any of the terms“comprising,” “consisting essentially of,” and “consisting of” may bereplaced with either of the other two terms. The terms and expressionswhich have been employed are used as terms of description and not oflimitation, and use of such terms and expressions do not exclude anyequivalents of the features shown and described or portions thereof, andvarious modifications are possible within the scope of the inventionclaimed. The term “a” or “an” can refer to one of or a plurality of theelements it modifies (e.g., “a reagent” can mean one or more reagents)unless it is contextually clear either one of the elements or more thanone of the elements is described. The term “about” as used herein refersto a value within 10% of the underlying parameter (i.e., plus or minus10%), and use of the term “about” at the beginning of a string of valuesmodifies each of the values (i.e., “about 1, 2 and 3” is about 1, about2 and about 3). For example, a weight of “about 100 grams” can includeweights between 90 grams and 110 grams. Further, when a listing ofvalues is described herein (e.g., about 50%, 60%, 70%, 80%, 85% or 86%)the listing includes all intermediate values thereof (e.g., 54%, 85.4%).Thus, it should be understood that although the present invention hasbeen specifically disclosed by representative embodiments and optionalfeatures, modification and variation of the concepts herein disclosedmay be resorted to by those skilled in the art, and such modificationsand variations are considered within the scope of this invention.

Certain embodiments of the invention are set forth in the claims thatfollow.

1. A method for detecting the presence or absence of a plurality oftarget alleles at a plurality of single nucleotide polymorphic loci in asample, wherein the sample contains nucleic acid, comprising thefollowing steps: a) cleaving nucleic acid comprising a plurality ofnon-target alleles at the plurality of single nucleotide polymorphicloci with Tsp509I restriction enzyme that recognizes the non-targetalleles, wherein the plurality of single nucleotide polymorphic loci areat positions selected from the group consisting of: rs11835780 rs748773rs7323716 rs4311632 rs6488494 rs9652080 rs13110085 rs1363267 rs10785736rs13269702 rs10840805 rs7356482 rs1797700 rs2723307 rs7831906 rs2993531rs4764597 rs9818611 rs1885121 rs4589569 rs12903747 rs1372688 rs2820107rs7320201 rs1904161 rs6766358 rs4869315 rs1720839 rs12675087 rs3913810rs10901705 rs7689368 rs6542638 rs6582294 rs725849 rs12450474 rs7144509rs7900002 rs1346718 rs10234234 rs910500 rs1503660 rs8016543 rs4489023rs2007475 rs11221881 rs1916803 rs683262 rs13155942 rs10260483 rs10110766rs494220 rs4488809 rs1041409 rs3912319 rs4533845 rs11099210 rs9428474rs3816551 rs10754776 rs9929404 rs6556642 rs4130306 rs7294836 rs7205009rs2734574 rs4673821 rs12674093 rs1401454 rs614004 rs2322301 rs9285190rs1444647 rs7741525 rs179596 rs7818415 rs17074340 rs331893 rs12007rs2462049 rs9787011 rs9356029 rs10806232 rs273172 rs6569474 rs11105611rs9989393 rs10898954 rs12107918 rs1342995 rs6043856 rs664358 rs263025rs1593443 and rs6142841;

b) amplifying uncleaved nucleic acid but not cleaved nucleic acid; andc) analyzing the amplification products of step b) to determine thepresence or absence of a plurality of target alleles, wherein the targetalleles are of paternal origin and the non-target alleles are ofmaternal origin.
 2. The method of claim 1, wherein the sample is from apregnant female or a female suspected of being pregnant.
 3. The methodof claim 1, wherein 20 or more of the single nucleotide polymorphic lociare assayed.
 4. The method of claim 1, wherein the target alleleconcentration is 10% or less of total nucleic acid concentration priorto cleaving and amplifying the nucleic acid.